TW201416361A - Inhibitors of Bruton's tyrosine kinase - Google Patents

Abstract

This application discloses compounds according to generic Formula I: wherein all variables are defined as described herein, which inhibit Btk. The compounds disclosed herein are useful to modulate the activity of Btk and treat diseases associated with excessive Btk activity. The compounds are further useful to treat inflammatory and auto immune diseases associated with aberrant B-cell proliferation such as rheumatoid arthritis. Also disclosed are compositions containing compounds of Formula I and at least one carrier, diluent or excipient.

Description

Bruton's tyrosine kinase inhibitor

The present invention relates to the use of novel compounds which inhibit Btk and which are useful in the treatment of autoimmune and inflammatory diseases caused by abnormal B cell activation.

Protein kinases constitute one of the largest family of enzymes in humans and regulate many different signaling processes by adding phosphate groups to proteins (T. Hunter, Cell 1987 50:823-829). In particular, tyrosine kinase phosphorylates proteins on the phenolic portion of tyrosine residues. The tyrosine kinase family includes members that control cell growth, migration, and differentiation. Abnormal kinase activity is associated with a variety of human diseases, including cancer, autoimmune diseases, and inflammatory diseases. Because protein kinases are one of the key regulators of cell signaling, they provide a target for small molecule kinase inhibitors to regulate cellular function and thus form good drug design targets. In addition to treating kinase-mediated disease processes, selective and potent inhibitors of kinase activity are also useful for studying cellular signaling processes and identifying other cellular targets of therapeutic importance.

There is sufficient evidence that B cells play a key role in the pathogenesis of autoimmune and/or inflammatory diseases. Protein-based therapeutics that deplete B cells, such as Rituxan, are effective against inflammatory diseases caused by autoantibodies, such as rheumatoid arthritis (Rastetter et al, Annu Rev Med 2004 55:477). Thus, inhibitors of protein kinases that play a role in B cell activation should be therapeutic agents suitable for B cell mediated disease pathologies, such as the production of autoantibodies.

A series of B cell responses can be controlled via B cell receptor (BCR) signaling, including proliferation and differentiation into cells that produce mature antibodies. BCR is a key regulatory point for B cell activity, and abnormal signaling leads to dysregulation of B cell proliferation and formation of pathogenic autoantibodies, resulting in a variety of autoimmune and/or inflammatory diseases. Bruton's Tyrosine Kinase (Btk) is a non-BCR-associated kinase that is closest to the membrane and is immediately downstream of the BCR. The lack of Btk has been shown to block BCR signaling, and thus inhibition of Btk may be a suitable therapeutic approach to block B cell mediated disease processes.

Btk is a member of the Tec family of tyrosine kinases and has been shown to be a key regulator of early B cell development and activation and survival of mature B cells (Khan et al, Immunity 1995 3: 283; Ellmeier et al., J. Exp. Med .2000 192:1611). Human Btk mutations cause disease X-linked gamma globulinemia (XLA) ( Resen et al., New Eng. J. Med. 1995 333:431 and Lindvall et al., Immunol. Rev. 2005 203:200) . These patients have immune dysfunction and show abnormal B cell maturation, decreased immunoglobulin and peripheral B cell content, reduced immune response unrelated to T cells, and reduced calcium movement following BCR stimulation.

Btk-deficient mouse models have also provided evidence of the role of Btk in autoimmune and inflammatory diseases. In a preclinical murine model of systemic lupus erythematosus (SLE), Btk-deficient mice showed a significant improvement in disease progression. Furthermore, Btk-deficient mice are resistant to collagen-induced arthritis (Jansson and Holmdahl Clin . Exp. Immunol. 1993 94: 459). Selective Btk inhibitors have shown dose-dependent efficacy in a mouse arthritis model (Z. Pan et al, Chem . Med Chem. 2007 2: 58-61).

Btk is also expressed by cells other than B cells that may be involved in the disease process. For example, Btk is expressed by mast cells and Btk-deficient bone marrow-derived mast cells show antigen-induced degranulation damage (Iwaki et al, J. Biol. Chem. 2005 280: 40261). This shows that Btk can be used to treat pathological mast cell responses such as allergies and asthma. Monocytes from XLA patients, in which Btk activity is absent, also showed a decrease in TNF alpha production following stimulation (Horwood et al, J Exp Med 197: 1603, 2003). Thus, small molecule Btk inhibitors can modulate TNF[alpha] mediated inflammation. Furthermore, Btk has been reported to play a role in apoptosis (Islam and Smith Immunol. Rev. 2000 178:49,) and therefore Btk inhibitors are indicated for the treatment of certain B cell lymphomas and leukemias (Feldhahn et al., J. Exp .Med .2005 201:1837).

The present application provides a Btk inhibitor compound of Formula I, methods of use thereof, as described below: This application provides a compound of Formula I,

among them: Is a single bond or a double bond; X is CH, CH ₂ or N; R is H, -R ¹ , -R ¹ -R ² -R ³ , -R ¹ -R ³ or -R ² -R ³ ; R ¹ An aryl group, a heteroaryl group, a bicyclic heteroaryl group, a cycloalkyl group or a heterocycloalkyl group, each of which may optionally be subjected to one or more lower alkyl, hydroxy, hydroxy lower alkyl, lower carbon numbers Alkoxy, halo, nitro, amine, decyl, cyano, pendant or lower haloalkyl; R ² is -C(=O), -C(=O)O, -C(=O)NR ^2' , -NHC(=O)O, -C(R ^2' ) ₂ , -O, -S, -C(=NH)NR ^2' or -S(=O) ₂ Each R ^{2 ' is} independently H or a lower alkyl group; R ³ is H or R ⁴ ; R ⁴ is a lower alkyl group, a lower carbon haloalkyl group, a lower alkoxy group, an amine group, Lower alkylalkylamino, cycloalkylamino, lower dialkylamino, aryl, arylalkyl, alkylaryl, heteroaryl, alkylheteroaryl, heteroarylalkane Base, cycloalkyl, alkylcycloalkyl, cycloalkylalkyl, heterocycloalkyl, alkylheterocycloalkyl, heterocycloalkylalkyl, bicyclic cycloalkyl, bicycloheterocycloalkyl, spiro ring An alkyl group, a spiroheterocycloalkyl group or a bicyclospirocycloalkyl group, each of which is optionally one or more C, alkyl, halo, lower alkylamino, lower dialkylamino, hydroxy, hydroxy lower alkyl, lower alkoxy, lower alkyl alkoxy, halo , nitro, amine, decyl, fluorenyl, cyano, pendant oxy, sulfonyl, lower alkyl sulfonyl, fluorenyl, hydroxyamino, carboxy, amine carbyl, amine a formate group, a halogen lower alkoxy group, a heterocycloalkyl group or a halogen lower alkyl group, wherein two lower alkyl groups may together form a ring; Y ⁴ is Y ^4a , Y ^4b , Y ^4c or Y ^4d ; Y ^4a is H or halogen; Y ^4b is a lower alkyl group, which is optionally selected from one or more selected from the group consisting of a lower halogen haloalkyl group, a halogen group, a hydroxyl group, an amine group, a cyano group and a lower group. Substituted by a substituent of the group of carbon number alkoxy groups; Y ^4c is a lower carbon cycloalkyl group, which is optionally selected from one or more selected from the group consisting of a lower alkyl group, a lower carbon haloalkyl group, a halogen group, a hydroxyl group, Substituted with a group of amine, cyano and lower alkoxy groups; and Y ^4d is an amine group, optionally via one or more lower alkyl, alkoxy lower alkyl or Hydroxylower alkyl substitution; or its medicinal Acceptable salt.

The application provides a method of treating an inflammatory and/or autoimmune condition comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a pharmaceutical composition comprising a mixture of a compound of formula I and at least one pharmaceutically acceptable carrier, excipient or diluent.

definition

The phrase "a", as used herein, refers to one or more such entities; for example, a compound refers to one or more compounds or at least one compound. Therefore, the terms "a", "one or more" and "at least one" are used interchangeably herein.

The phrase "as defined above" refers to the broadest definition of each group as provided in the Summary of the Invention or in the broadest scope of application. In all other embodiments provided below, the substituents that may be present in various embodiments and which are not explicitly defined retain the broadest definitions provided in the Summary of the Invention.

As used in this specification, the term "comprising" should be interpreted as having an open meaning, whether in the context of a patent's transitional phrase or in its subject. That is, the term should be interpreted as synonymous with the words "at least have" or "at least include". The term "comprising" when used in the context of a method means that the method includes at least the steps recited, but may include additional steps. The term "comprising" when used in the context of a compound or composition means that the compound or composition includes at least the features or components, but may also include other features or components.

The word "or" as used herein is used in the sense of "including" or "exclusive" of "and" or "exclusive".

The term "independent" is used herein to mean that a variable is used in either condition, regardless of the presence or absence of a variable having the same or different definitions in the same compound. Thus, in a compound where R" occurs twice and is defined as "independent carbon or nitrogen", both R" can be carbon, both R" can be nitrogen or one R" can be carbon and the other nitrogen .

When any variable occurs more than once in any part or chemical formula depicting and describing a compound employed or claimed by the present invention, its definition at each occurrence is independent of the definition at the time of its other occurrence. Also, it is permissible to take only when such combinations produce stable compounds. A combination of a base and/or a variable part.

The symbol "*" at the end of the key or drawn through a button Each refers to the point of attachment of a functional group or other chemical moiety to the rest of the molecule to which the functional group or other chemical moiety belongs. So, for example: MeC(=O)OR ⁴ where R ⁴ = or MeC(=O)O .

The keys drawn into the ring system (as opposed to being joined at the respective vertices) indicate that the bond can be attached to any of the appropriate ring atoms.

The term "optionally" or "as appropriate" as used herein means that the subsequently described event or circumstance may (but is not required to) occur and that the description includes the occurrence of the event or circumstance and that the event and event or circumstance has not occurred. The situation. For example, "substituted as appropriate" means that a portion substituted as the case may have a hydrogen atom or a substituent.

The phrase "optionally selected key" means that the key may or may not exist, and the description includes a single key, a double key or a triple key. If a substituent is designated as "one bond" or "absent", the atom to which the substituent is attached is directly linked.

The term "about" as used herein means approximation, in scope, roughly or left and right. When the term "about" is used in conjunction with a range of values, the range can be revised by expanding the bounds of the set number of values up and down. Generally, the term "about" as used herein is used to modify the value above and below the 20% deviation.

Certain compounds of formula I can exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Proton-shifting tautomers arise from the migration of hydrogen atoms covalently bonded between two atoms. Tautomers generally assume an equilibrium state and attempts to separate individual tautomers typically result in a mixture whose chemical and physical properties conform to a mixture of compounds. The equilibrium position depends on the chemical characteristics within the molecule. For example, in many aliphatic aldehydes and aliphatic ketones (such as acetaldehyde), the predominant form is the keto form; and in phenol, the predominant form is the enol form. Common proton transfer tautomers include ketone/enol (-C(=O)-CH- -C(-OH)=CH-), decylamine/imidic acid (-C(=O)-NH- -C(-OH)=N-) and 脒(-C(=NR)-NH- -C(-NHR)=N-) tautomer. The latter two are especially common in heteroaryl and heterocycles and the invention encompasses all tautomeric forms of such compounds.

Unless otherwise defined, the technical terms used herein have the meaning commonly understood by those of ordinary skill in the art. Various methods and materials known to those skilled in the art are mentioned herein. Standard references describing the general principles of pharmacology include Goodman and Gilman, The Pharmacological Basis of Therapeutics , 10th Edition, McGraw Hill Companies Inc., New York (2001). Any suitable materials and/or methods known to those skilled in the art can be used in the practice of the present invention. However, preferred materials and methods are described. Unless otherwise indicated, the materials, reagents, and analogs referred to in the following description and examples are available from commercial sources.

The definitions described herein may be appended to form chemically relevant combinations such as "heteroalkylaryl", "haloalkylheteroaryl", "arylalkylheterocyclyl", "alkylcarbonyl", "alkane" Oxyalkyl groups and the like. When the term "alkyl" is used as the suffix of another term, as in "phenylalkyl" or "hydroxyalkyl", this is intended to mean that the alkyl group as defined above is selected from 1 to 2 Substituted by a substituent of a specifically named group. Thus, for example, "phenylalkyl" refers to an alkyl group having from 1 to 2 phenyl substituents and thus includes benzyl, phenethyl and biphenyl. "Alkylaminoalkyl" is an alkyl group having one to two alkylamino substituents. "Hydroxyalkyl" includes 2-hydroxyethyl, 2-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 2,3-dihydroxybutyl, 2- (hydroxymethyl), 3-hydroxypropyl, and the like. Thus, the term "hydroxyalkyl" as used herein, is used to define one of the heteroalkyl groups defined below. The term -(aryl)alkyl means an unsubstituted alkyl or aralkyl group. The term (hetero)aryl refers to an aryl or heteroaryl group.

The term "spirocycloalkyl" as used herein means spirocycloalkyl, such as spiro[3.3]heptane. The term spiroheterocycloalkyl as used herein means spirocycloheteroalkyl, such as 2,6-diazaspiro[3.3]heptane.

The term "mercapto" as used herein denotes a radical of the formula -C(=O)R, wherein R is hydrogen or a lower alkyl group as defined herein. The term "alkylcarbonyl," as used herein, refers to a radical of the formula C(=O)R, wherein R is alkyl as defined herein. The term C _1-6 fluorenyl refers to a group -C(=O)R having from 1 to 6 carbon atoms. The term "arylcarbonyl," as used herein, means a radical of the formula C(=O)R, wherein R is aryl; as used herein, the term "benzimidyl" is an arylcarbonyl group wherein R is phenyl. "."

The term "ester" as used herein denotes a radical of the formula -C(=O)OR, wherein R is lower alkyl as defined herein.

The term "alkyl" as used herein denotes an unbranched or branched, saturated, monovalent hydrocarbon residue containing from 1 to 10 carbon atoms. The term "lower alkyl" means a straight or branched hydrocarbon residue having from 1 to 6 carbon atoms. "C _{1 -10} alkyl" as used herein means an alkyl group containing from 1 to 10 carbons. Examples of alkyl groups include, but are not limited to, lower alkyl groups including methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl or pentyl, isopentyl , neopentyl, hexyl, heptyl and octyl.

When the term "alkyl" is used as the suffix of another term, as in "phenylalkyl" or "hydroxyalkyl", this is intended to mean that the alkyl group as defined above is selected from 1 to 2 Substituted by a substituent of a specifically named group. Thus, for example, "phenylalkyl" refers to the group R'R"-, wherein R' is phenyl, and R" is alkyl as defined herein, provided that the phenylalkyl moiety is attached The point will be on the alkyl group. Examples of arylalkyl groups include, but are not limited to, benzyl, phenethyl, 3-phenylpropyl. The terms "arylalkyl" or "aralkyl" are interpreted similarly, except that R' is aryl. The term "(hetero)arylalkyl" or "(hetero)aralkyl" is interpreted similarly, except that R' is optionally aryl or heteroaryl.

The term "haloalkyl" or "halo-lower alkyl" or "lower haloalkyl" refers to a straight or branched hydrocarbon residue containing from 1 to 6 carbon atoms, one or more of which The carbon atom is substituted with one or more halogen atoms.

Unless otherwise indicated, as used herein, the term "alkylene" denotes a divalent from 1 to 10 carbon atoms of straight-chain saturated hydrocarbon group (e.g., (CH _{2) n)} or a branched chain having 2 to 10 carbon atoms A saturated divalent hydrocarbon group (for example, -CHMe- or -CH ₂ CH( i -Pr)CH ₂ -). Except in the case of methylene groups, the open price of an alkylene group is not bonded to the same atom. Examples of alkylene groups include, but are not limited to, methylene, ethyl, propyl, 2-methyl-propyl, 1,1-dimethyl-ethyl, butyl, 2- Ethyl butyl.

The term "alkoxy" as used herein means -O-alkyl, wherein alkyl is as defined above, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, Isobutoxy, tert-butoxy, pentyloxy, hexyloxy, including isomers thereof. As used herein, "lower alkoxy" means an alkoxy group having a "lower alkyl group" as defined above. "C _{1 -10} alkoxy" as used herein means an -O-alkyl group wherein the alkyl group is C _1-10 .

The term "PCy ₃ " refers to a phosphine that is trisubstituted with three cyclic moieties.

The term "haloalkoxy" or "halo-lower alkoxy" or "lower haloalkoxy" refers to a low carbon number in which one or more carbon atoms are replaced by one or more halogen atoms. Alkoxy.

The term "hydroxyalkyl" as used herein denotes that one to three hydrogen atoms on different carbon atoms in an alkyl group, as defined herein, are replaced by a hydroxy group.

The terms "alkylsulfonyl" and "arylsulfonyl" as used herein mean a radical of the formula -S(=O) ₂ R wherein R is alkyl or aryl, respectively, and alkyl and aryl The base is as defined herein. The term "heteroalkylsulfonyl" as used herein denotes a radical of the formula -S(=O) ₂ R, wherein R is "heteroalkyl" as defined herein.

The terms "alkylsulfonylamino" and "arylsulfonylamino" as used herein mean a radical of the formula -NR'S(=O) ₂ R wherein R is alkyl or aryl, respectively, and R' is Hydrogen or C _1-3 alkyl and alkyl and aryl are as defined herein.

The term "cycloalkyl" as used herein refers to a saturated carbocyclic ring containing from 3 to 8 carbon atoms, ie, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. As used herein, "C _3-7 cycloalkyl" refers to a cycloalkyl group containing from 3 to 7 carbons in the carbocyclic ring.

The term carboxy-alkyl as used herein refers to an alkyl moiety in which one hydrogen atom is replaced by a carboxy group, provided that the point of attachment of the heteroalkyl group passes through the carbon atom. The term "carboxy" refers to the -CO ₂ H moiety.

The term "heteroaryl" or "heteroaromatic" as used herein, means having from 5 to 12 ring atoms and having at least one aromatic or partially unsaturated ring (each ring containing from 4 to 8 atoms), combined with Or a plurality of N, O or S heteroatoms, the remaining ring atoms being a monocyclic or bicyclic group of carbon, provided that the point of attachment of the heteroaryl group will be on the aromatic or partially unsaturated ring. As is well known to those skilled in the art, the heteroaryl ring has fewer aromatic features than its all carbon counterpart. Thus, for the present invention, the heteroaryl group only needs to have a certain degree of aromatic character. Examples of heteroaryl moieties include monocyclic aromatic heterocycles having 5 to 6 ring atoms and 1 to 3 heteroatoms including, but not limited to, pyridinyl, pyrimidinyl, pyrazinyl, oxazinyl, pyrrole Base, pyrazolyl, imidazolyl, oxazolyl, 4,5-dihydro-oxazolyl, 5,6-dihydro-4H-[1,3]oxazolyl, isoxazole, thiazole, isothiazole , a triazoline, a thiadiazole, and an oxadiazoline, optionally substituted by one or more, preferably 1 or 2 substituents selected from the group consisting of hydroxyl, cyano, alkyl, alkoxy, Thio group, lower carbon number haloalkoxy group, alkylthio group, halo group, lower carbon haloalkyl group, alkyl sulfinyl group, alkyl sulfonyl group, halogen, amine group, alkyl amine group , dialkylamino, aminoalkyl, alkylaminoalkyl and dialkylaminoalkyl, nitro, alkoxycarbonyl and aminecarbamyl, alkylaminecarbamyl, dialkylamine A decyl group, an arylamine carbhydryl group, an alkylcarbonylamino group, and an arylcarbonylamino group. Examples of bicyclic moieties include, but are not limited to, quinolyl, isoquinolyl, benzofuranyl, benzothienyl, benzoxazole, benzisoxazole, benzothiazole, Pyridyl, 5,6,7,8-tetrahydro-[1,6] Pyridyl and benzisothiazole. The bicyclic moiety may be substituted on either ring as appropriate, however the attachment point is on a ring containing a hetero atom.

The terms "heterocyclyl", "heterocycloalkyl" or "heterocycle" as used herein, unless otherwise indicated, mean one or more rings, preferably 1 to 2 rings (including a spiro ring system). a composition, each ring having 3 to 8 atoms, combined with one or more ring heteroatoms (selected from N, O or S(O) _0-2 ), a monovalent saturated cyclic group, which may optionally be one or a plurality of, preferably one or two, substituents selected from the group consisting of a hydroxyl group, a pendant oxy group, a cyano group, a lower alkyl group, a lower alkoxy group, a lower number of haloalkoxy groups, Alkylthio, halo, lower halohaloalkyl, hydroxyalkyl, nitro, alkoxycarbonyl, amine, alkylamino, alkylsulfonyl, arylsulfonyl, alkylamino Sulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamine Base and its ionic form. Examples of heterocyclic groups include, but are not limited to, morpholinyl, piperazinyl, piperidinyl, azetidinyl, pyrrolidinyl, hexahydroazepine, oxetanyl, tetrahydrofuranyl , tetrahydrothiophenyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, tetrahydropyranyl, thiomorpholinyl, Pyridyl and imidazolinyl, and their ionic forms. Examples may also be bicyclic, such as 3,8-diaza-bicyclo[3.2.1]octane, 2,5-diaza-bicyclo[2.2.2]octane or octahydro-pyrazine[2, 1-c][1,4]oxazine.

Btk inhibitor

This application provides a compound of formula I,

among them: Is a single bond or a double bond; X is CH, CH ₂ or N; R is H, -R ¹ , -R ¹ -R ² -R ³ , -R ¹ -R ³ or -R ² -R ³ ; R ¹ An aryl group, a heteroaryl group, a bicyclic heteroaryl group, a cycloalkyl group or a heterocycloalkyl group, each of which may optionally be subjected to one or more lower alkyl, hydroxy, hydroxy lower alkyl, lower carbon numbers Alkoxy, halo, nitro, amine, decyl, cyano, pendant or lower haloalkyl; R ² is -C(=O), -C(=O)O, -C(=O)NR ^2' , -NHC(=O)O, -C(R ^2' ) ₂ , -O, -S, -C(=NH)NR ^2' or -S(=O) ₂ Each R ^{2 ' is} independently H or a lower alkyl group; R ³ is H or R ⁴ ; R ⁴ is a lower alkyl group, a lower carbon haloalkyl group, a lower alkoxy group, an amine group, Lower alkylalkylamino, cycloalkylamino, lower dialkylamino, aryl, arylalkyl, alkylaryl, heteroaryl, alkylheteroaryl, heteroarylalkane Base, cycloalkyl, alkylcycloalkyl, cycloalkylalkyl, heterocycloalkyl, alkylheterocycloalkyl, heterocycloalkylalkyl, bicyclic cycloalkyl, bicycloheterocycloalkyl, spiro ring An alkyl group, a spiroheterocycloalkyl group or a bicyclospirocycloalkyl group, each of which is optionally one or more low C, alkyl, halo, lower alkylamino, lower dialkylamino, hydroxy, hydroxy lower alkyl, lower alkoxy, lower alkyl alkoxy, halo , nitro, amine, decyl, fluorenyl, cyano, pendant oxy, sulfonyl, lower alkyl sulfonyl, fluorenyl, hydroxyamino, carboxy, amine carbyl, amine a formate group, a halogen lower alkoxy group, a heterocycloalkyl group or a halogen lower alkyl group, wherein two lower alkyl groups may together form a ring; Y ⁴ is Y ^4a , Y ^4b , Y ^4c or Y ^4d ; Y ^4a is H or halogen; Y ^4b is a lower alkyl group, which is optionally selected from one or more selected from the group consisting of a lower halogen haloalkyl group, a halogen group, a hydroxyl group, an amine group, a cyano group and a lower group. Substituted by a substituent of the group of carbon number alkoxy groups; Y ^4c is a lower carbon cycloalkyl group, which is optionally selected from one or more selected from the group consisting of a lower alkyl group, a lower carbon haloalkyl group, a halogen group, a hydroxyl group, Substituted with a group of amine, cyano and lower alkoxy groups; and Y ^4d is an amine group, optionally via one or more lower alkyl, alkoxy lower alkyl or Hydroxylower alkyl substitution; or its medicinal Acceptable salt thereof.

Furthermore, it is to be understood that each of the embodiments disclosed herein in connection with a particular residue R, R ¹ , R ² , R ³ , R ⁴ , X, R ^2, and Y ⁴ can be associated with any other as disclosed herein. Combinations of examples relating to another residue R, R ¹ , R ² , R ³ , R ⁴ , X, R ^2, and Y ⁴ .

The application provides a compound of formula I, wherein Is a double bond; and X is N.

The application provides a compound of formula I, wherein Is a single bond; and X is CH ₂ .

The application provides a compound of formula I, wherein Is a double bond; X is N; R is -R ¹ -R ² -R ³ ; R ¹ is pyridyl; R ² is -S(=O) ₂ ; R ³ is R ⁴ ; and R ⁴ is a low carbon number alkyl.

The application provides a compound of formula I, wherein Is a double bond; X is N; R is -R ¹ -R ² -R ³ ; R ¹ is pyridyl; R ² is -C(CH ₃ ) ₂ ; R ³ is R ⁴ ; and R ⁴ is a low carbon number An alkylamino group, a lower dialkylamino group or a heterocycloalkyl group optionally substituted with one or more lower alkyl groups.

The application provides a compound of formula I, wherein Is a double bond; X is N; R is -R ¹ -R ² -R ³ ; R ¹ is phenyl or pyridyl; R ² is -C(=O); R ³ is R ⁴ ; and R ⁴ is it? A phenyl or piperazine, optionally substituted with one or more lower alkyl groups.

The application provides a compound of formula I, wherein Is a double bond; X is N; and Y ⁴ is a third butyl group.

The application provides a compound of formula I, wherein Is a double bond; X is CH; and Y ⁴ is

Wherein Y ⁵ and Y ^{6 are} independently H, a lower alkyl group or a lower carbon haloalkyl group.

The application provides a compound of formula I, wherein Is a double bond; X is N; and Y ⁴ is

Wherein Y ⁵ is H, halogen, lower alkyl or lower haloalkyl.

The application provides a compound of formula I, wherein Is a double bond; X is N; and Y ⁴ is

Wherein Y ⁵ and Y ^{6 are} independently H or a lower alkyl group.

The application provides a compound of formula I, wherein Is a double bond; X is N; Y ⁴ is a third butyl group; R is -R ¹ -R ³ ; R ¹ is pyridyl or pyrazolopyrazine; R ³ is R ⁴ ; and R ⁴ is optionally Substituted lower alkyl, heterocycloalkyl or alkylheterocycloalkyl.

The application provides a compound of formula I, wherein Is a double bond; X is N; Y ⁴ is a third butyl group; R is -R ¹ -R ² -R ³ ; R ¹ is a pyridyl group; R ² is -C(CH ₃ ) ₂ ; R ³ is R ⁴ And R ⁴ is a lower alkylamino group, a lower dialkylamino group or a heterocycloalkyl group optionally substituted with one or more lower alkyl groups.

The application provides a compound of formula I, wherein Is a double bond; X is N; Y ⁴ is a third butyl group; R is -R ¹ -R ² -R ³ ; R ¹ is a pyridyl group; R ² is -C(=O); R ³ is R ⁴ ; And R ⁴ is optionally substituted heterocycloalkyl or bicyclospirocycloalkyl.

The application provides a compound of formula I, wherein Is a double bond; X is N; Y ⁴ is a third butyl group; R is -R ¹ -R ² -R ³ ; R ¹ is a pyridyl group; R ² is -C(=O); R ³ is R ⁴ ; And R ⁴ is a morpholine or piperazine which is optionally substituted.

The application provides a compound of formula I selected from the group consisting of 2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-B) Oxy)-pyridin-2-ylamino]-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-6 - tert-butyl-2H-phthalazin-1-one; 6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1' , 2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazine 3-yl]-phenyl}-2H-phthalazin-1-one; 2-{3-[5-(5-azetidin-1-ylmethyl-1-methyl-1H-pyridyl) Zin-3-ylamino)-1-yl 6-o-oxy-1,6-dihydro-pyridazin-3-yl]-2-hydroxymethyl-phenyl}-6-tert-butyl-2H-phthalazin-1-one; -T-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((S)-1-methyl-pyrrolidin-2-yl)-pyridine-2- Amino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one; 6-t-butyl-2-(2) -hydroxymethyl-3-{1-methyl-5-[5-((S)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-6-sideoxy -1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one; 6-t-butyl-2-(2-hydroxymethyl-3-{1- Methyl-5-[5-((R)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-6-oxo-oxy-1,6-dihydro-pyridazine -3-yl}-phenyl)-2H-phthalazin-1-one; 2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1' , 2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazine 3-yl]-phenyl}-1-o-oxy-1,2-dihydro-pyridazin-6-yl)-2-methyl-propionitrile; 6-t-butyl-2-{2 -hydroxymethyl-3-[1-methyl-5-(5-methyl-4,5,6,7-tetrahydro-pyrazolo[1,5-a]pyrazin-2-ylamino 6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-2H-pyridazin-1-one; 6-t-butyl-2-{3-[5 -(1'-ethyl-1', 2', 3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-1-methyl-6-o-oxy-1,6-dihydro-indole Pyrazin-3-yl]-2-hydroxymethyl-phenyl}-2H-phthalazin-1-one; 6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl- 5-[5-(morpholine-4-carbonyl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H- Pyridazin-1-one; 2-[2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridine-2 -ylamino]-1-methyl-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-1-yloxy-1, 2-dihydro-pyridazin-6-yl]-2-methyl-propionitrile; 6-Terbutyl-2-[2-hydroxymethyl-3-(5-{5-[2-(3-hydroxy-propylamino)-1,1-dimethyl-ethoxy] -pyridin-2-ylamino}-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl)-phenyl]-2H-pyridazin-1-one; -T-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(1'-oxetan-3-yl-1',2',3',4' , 5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl} -2H-phthalazin-1-one; 6-t-butyl-2-[3-(5-{5-[2-(3,3-difluoro-azetidin-1-yl)- 1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxirane-1,6-dihydro-pyridazin-3-yl)-2- Hydroxymethyl-phenyl]-2H-phthalazin-1-one; and 2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1', 2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazine- 3-yl]-phenyl}-1-sidedoxy-1,2,3,4-tetrahydro-isoquinolin-6-yl)-2-methyl-propanenitrile.

The application provides a method of treating an inflammatory or autoimmune condition comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a method of treating rheumatoid arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a pharmaceutical composition comprising a compound of formula I.

The application provides a pharmaceutical composition comprising a mixture of a compound of formula I and at least one pharmaceutically acceptable carrier, excipient or diluent.

The application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of an inflammatory condition.

This application provides the use of a compound of formula I for the manufacture of a self-immunity The agent of the epidemic.

The application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of rheumatoid arthritis.

The application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of asthma.

The application provides the use of a compound as described above for the treatment of an inflammatory or autoimmune disorder.

The application provides the use of a compound as described above for the treatment of rheumatoid arthritis.

The application provides the use of a compound as described above for the treatment of asthma.

The application provides a compound as described above for use in the treatment of an inflammatory or autoimmune disorder.

The application provides a compound as described above for the treatment of rheumatoid arthritis.

The application provides a compound as described above for use in the treatment of asthma.

The application provides a compound, method or composition as described herein.

The application provides a method of treating an inflammatory or autoimmune disorder comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I'.

The application provides a method of treating arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I'.

The application provides a method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I'.

The application provides a method of inhibiting B cell proliferation comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I'.

The application provides a method of inhibiting Btk activity comprising administering a Btk inhibitor compound having any one of Formula I', wherein the Btk inhibitor compound exhibits 50 micromoles or 50 micrograms in an in vitro biochemical assay of Btk activity. The IC ₅₀ below Moher.

In a variant of the above method, Btk inhibitor compound exhibits nemorubicin ear or ear 100 of the IC ₅₀ 100 nemorubicin less in an in vitro biochemical assay of Btk activity in the.

In another variant of the above method, the compound exhibits 10 nemorubicin nemorubicin ear or ears 10 of the IC ₅₀ in an in vitro biochemical assay of Btk activity in the.

The present application provides a method of treating an inflammatory condition comprising a combination of a therapeutically effective amount of an anti-inflammatory compound and a Btk inhibitor compound of Formula I, co-administered to a patient in need thereof.

The present application provides a method of treating arthritis comprising a combination of a therapeutically effective amount of an anti-inflammatory compound and a Btk inhibitor compound of Formula I, co-administered to a patient in need thereof.

The present application provides a method of treating lymphoma or BCR-ABL1 ⁺ leukemia cells by administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I.

The application provides a pharmaceutical composition comprising a mixture of a Btk inhibitor compound of Formula I and at least one pharmaceutically acceptable carrier, excipient or diluent.

The application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of an inflammatory condition.

The application provides the use of a compound of formula I' for the manufacture of a medicament for the treatment of an autoimmune disorder.

The application provides a compound, method or composition as described herein.

Compound and preparation method

Examples of representative compounds encompassed within the scope of the invention and which are within the scope of the invention are provided in the following table. The examples are provided and the subsequent methods of preparation are provided to enable those skilled in the art to understand and practice the invention. It is not intended to limit the scope of the invention, but is merely illustrative and representative of the invention.

In general, the nomenclature used in this application is based on the version 4.0 AUTONOM ^TM, i.e. for generating a Beilstein Institute computerized system of the IUPAC naming system. If there is a difference between the described structure and the name given by the structure, the structure described will control. Furthermore, if a stereochemistry of a structure or a portion of a structure is not indicated, for example, in bold or dashed lines, the structure or portions of the structure are understood to include all stereoisomers thereof.

Table I depicts examples of pyridazinone compounds of formula I:

General synthetic process

This application provides a method of preparing a compound of formula II wherein Y ₁ is Acid or pinacol An acid ester comprising the following steps:

The compound of formula I is heated to a temperature of from about 40 ° C to 150 ° C in the presence of a bis-pinacol borate, a palladium catalyst, a base and a phosphine.

The application provides the above process wherein the phosphine is PCy3, an alkyl monophosphine compound, an aryl monophosphine compound, an alkyl diphosphorus compound or an aryl diphosphine compound.

The present application provides the above process wherein the base is an inorganic base comprising potassium carbonate, cesium carbonate, potassium phosphate and potassium acetate, or an amine base including dicyclohexylamine and triethylamine.

The present application provides the above process, wherein the palladium catalyst is palladium (II) acetate or bis(dibenzylideneacetone)palladium (0), ginseng (diphenylmethyleneacetone) dipalladium (0) or the like.

This application provides a method of preparing a compound of Formula IV, which comprises the steps of:

a) a compound of formula II in the presence of a compound of formula III, a palladium catalyst, a base and a phosphine (wherein Y ₁ is Acid or pinacol The acid ester or a mixture of the two) is heated to about 40 ° C to 150 ° C; and b) with a base such as sodium hydroxide or hydroxide in a solvent such as methanol at a temperature between about room temperature and about 40 ° C. The product of step a) is treated with potassium or potassium carbonate or cesium carbonate.

The application provides the above process wherein the phosphine is PCy3, an alkyl monophosphine compound, an aryl monophosphine compound, an alkyl diphosphorus compound or an aryl diphosphine compound.

The present application provides the above process wherein the base is an inorganic base comprising potassium carbonate, cesium carbonate, potassium phosphate and potassium acetate, or an amine base including dicyclohexylamine and triethylamine.

The present application provides the above process, wherein the palladium catalyst is palladium (II) acetate or bis(dibenzylideneacetone)palladium (0), ginseng (diphenylmethyleneacetone) dipalladium (0) or the like.

This application provides a method of preparing a compound of Formula IV, which comprises the steps of:

a) heating the compound of formula I to a temperature of from about 40 ° C to 150 ° C in the presence of a bis-pinacol borate, a palladium catalyst, a base and a phosphine; b) in the presence of a compound of formula VII, a palladium catalyst, a base and a phosphine Step a) separation of the product with a compound of formula III to about 40 ° C to 150 ° C; and c) using a base (such as sodium hydroxide) in a solvent (such as methanol) at a temperature between about room temperature and about 40 ° C Or the product of step b) by treatment with potassium hydroxide or potassium carbonate or cesium carbonate.

This application provides a method of preparing a compound of Formula IV, which comprises the steps of:

a) a compound of formula II in the presence of a compound of formula V, a palladium catalyst, a base and a phosphine (wherein Y ₁ is Acid or pinacol An acid ester or a mixture of the two) is heated to a temperature of from about 40 ° C to 150 ° C; and b) a reducing agent such as sodium borohydride or a solvent thereof in a solvent such as methanol or a mixture of methanol and water at a temperature of about room temperature Analog)) The product of step a).

The application provides the above process wherein the phosphine is PCy3, an alkyl monophosphine compound, an aryl monophosphine compound, an alkyl diphosphorus compound or an aryl diphosphine compound.

The present application provides the above process wherein the base is an inorganic base comprising potassium carbonate, cesium carbonate, potassium phosphate and potassium acetate, or an amine base including dicyclohexylamine and triethylamine.

The present application provides the above process, wherein the palladium catalyst is palladium (II) acetate or bis(dibenzylideneacetone)palladium (0), ginseng (diphenylmethyleneacetone) dipalladium (0) or the like.

The application provides a method of preparing a compound of formula VII, which comprises the steps of:

a) a compound of formula II in the presence of a compound of formula VI, a palladium catalyst, a base and a phosphine (wherein Y ₁ is Acid or pinacol The acid ester or a mixture of the two) is heated to about 40 ° C to 150 ° C; and b) at a temperature of about room temperature in a solvent such as methanol or a mixture of methanol and water, with a reducing agent such as sodium borohydride or Its analog) treats the product of step a).

The application provides the above process wherein the phosphine is PCy3, an alkyl monophosphine compound, an aryl monophosphine compound, an alkyl diphosphorus compound or an aryl diphosphine compound.

The present application provides the above process wherein the base is an inorganic base comprising potassium carbonate, cesium carbonate, potassium phosphate and potassium acetate, or an amine base including dicyclohexylamine and triethylamine.

The present application provides the above process, wherein the palladium catalyst is palladium (II) acetate or bis(dibenzylideneacetone)palladium (0), ginseng (diphenylmethyleneacetone) dipalladium (0) or the like.

The present application provides a process for the preparation of a compound of formula VIII, wherein Y ₁ is Acid or pinacol An acid ester comprising the following steps:

The compound of formula V is heated in the presence of a bis-pinacol borate, a palladium catalyst, a base and a phosphine.

The application provides the above process wherein the phosphine is PCy3, an alkyl monophosphine compound, an aryl monophosphine compound, an alkyl diphosphorus compound or an aryl diphosphine compound.

The present application provides the above process wherein the base is an inorganic base comprising potassium carbonate, cesium carbonate, potassium phosphate and potassium acetate, or an amine base including dicyclohexylamine and triethylamine.

The present application provides the above process, wherein the palladium catalyst is palladium (II) acetate or bis(dibenzylideneacetone)palladium (0), ginseng (diphenylmethyleneacetone) dipalladium (0) or the like.

This application provides a method of preparing a compound of Formula IV, which comprises the steps of:

a) a compound of formula VIII (where Y ₁ is in the presence of a compound of formula I, a palladium catalyst, a base and a phosphine) Acid or pinacol An acid ester or a mixture of the two) is heated to a temperature of from about 40 ° C to 150 ° C; and b) a reducing agent such as sodium borohydride or a solvent thereof in a solvent such as methanol or a mixture of methanol and water at a temperature of about room temperature Analog)) The product of step a).

The application provides the above process wherein the phosphine is PCy3, an alkyl monophosphine compound, an aryl monophosphine compound, an alkyl diphosphorus compound or an aryl diphosphine compound.

The present application provides the above process wherein the base is an inorganic base comprising potassium carbonate, cesium carbonate, potassium phosphate and potassium acetate, or an amine base including dicyclohexylamine and triethylamine.

The present application provides the above process, wherein the palladium catalyst is palladium (II) acetate or bis(dibenzylideneacetone)palladium (0), ginseng (diphenylmethyleneacetone) dipalladium (0) or the like.

Pharmaceutical composition and administration

The compounds of the invention may be formulated in a variety of oral dosage forms and carriers. Oral administration can be in the form of lozenges, coated lozenges, dragees, hard gelatin capsules and soft gelatin capsules, solutions, emulsions, syrups or suspensions. The compounds of the invention are also effective when administered by other routes of administration, including continuous (intravenous infusion) topical parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include penetration) Reinforcing agent), buccal, nasal, inhalation and suppository administration. A preferred mode of administration is usually a convenient daily oral administration which is adjusted depending on the degree of the disease and the patient's response to the active ingredient.

The compounds of the present invention, as well as the pharmaceutically acceptable salts thereof, may be formulated in a pharmaceutical composition and unit dosage form together with one or more conventional excipients, carriers or diluents. The pharmaceutical compositions and unit dosage forms may contain conventional ingredients in the ordinary proportions, with or without other active compounds or ingredients, and the unit dosage form may contain the desired daily dosage range to be used Corresponding to any suitable effective amount of the active ingredient. The pharmaceutical composition may be used in the form of a solid such as a lozenge or a filled capsule; a semisolid; a powder; a sustained release formulation; or a liquid such as a solution, suspension, emulsion, elixirs or capsules for oral administration; or In the form of a suppository for rectal or vaginal administration; or in the form of a sterile injectable solution for parenteral use. A typical preparation contains from about 5% to about 95% active compound (w/w). The term "formulation" or "dosage form" is intended to include both solid and liquid formulations of the active compound and those skilled in the art will appreciate that the active ingredient may be present in different formulations depending on the target organ or Learning parameters.

The term "excipient" as used herein refers to a compound suitable for the preparation of a pharmaceutical composition, which is generally safe, non-toxic, and biologically and otherwise undesirable, and which includes veterinary medicine. Uses and excipients accepted for human medical use. The compounds of the present invention can be administered alone, but are usually administered in admixture with one or more appropriate pharmaceutical excipients, diluents or carriers selected according to the desired route of administration and standard pharmaceutical practice.

"Pharmaceutically acceptable" means that it is suitable for the preparation of a pharmaceutical composition which is generally safe, non-toxic and which is neither biologically undesirable nor otherwise undesirable and which is acceptable for veterinary and human medical use.

The "pharmaceutically acceptable salt" form of the active ingredient may also initially impart the desired pharmacokinetic properties which are not present in the non-salt form of the active ingredient and may even positively affect the pharmacodynamics of the active ingredient in the context of its therapeutic activity. "Pharmaceutically acceptable salt" of a phrase compound means a salt which is pharmaceutically acceptable and which possesses the desired pharmacological activity of the parent compound. The salts include: (1) an acid addition salt formed with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or an acid addition salt formed with an organic acid, organic Acids such as acetic acid, propionic acid, caproic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, hydroxysuccinic acid, maleic acid, fumaric acid, tartaric acid , citric acid, benzoic acid, 3-(4-hydroxybenzhydryl)benzoic acid, cinnamic acid, almond Acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluene Sulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butyl Acetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) when the acid proton present in the parent compound is Metal ions (such as alkali metal ions, alkaline earth ions or aluminum ions) replace the formed salt; or with organic bases (such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucosamine and similar bases) The salt formed during the position.

Solid form preparations include powders, lozenges, pills, capsules, cachets, suppositories, and dispersible granules. The solid carrier can be one or more substances which may also act as a diluent, a flavoring agent, a solubilizer, a lubricant, a suspending agent, a binder, a preservative, a tablet disintegrating agent or an encapsulating material. In powders, the carrier is usually a finely divided solid in admixture with the finely divided active component. In tablets, the active component is usually mixed in a suitable ratio with the carrier having the necessary adhesive strength and compressed into the desired shape and size. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, Low melting wax, cocoa butter and similar carriers. The solid form preparation may contain, in addition to the active ingredient, coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersing agents, thickening agents, solubilizing agents, and the like.

Liquid formulations are also suitable for oral administration, including emulsions, syrups, spirits, aqueous solutions and aqueous suspensions. Such formulations include solid form preparations which are intended to be converted to liquid form preparations prior to use. The emulsion may be prepared in a solution, such as an aqueous solution of propylene glycol, or may contain an emulsifier such as lecithin, sorbitan monooleate or gum arabic. The aqueous solution can be prepared by dissolving the active component in water and adding suitable color formers, flavoring agents, stabilizers, and thickening agents. Aqueous suspensions can be prepared by dispersing the finely divided active component in water in a viscous material such as natural or synthetic gum, resin, methylcellulose, sodium carboxymethylcellulose, and other well known suspending agents.

The compounds of the invention may be formulated for parenteral administration (for example by injection, such as bolus injection or continuous infusion), and may be presented in unit dosage form in ampoules, prefilled syringes, small volume infusions or with added antiseptic The agents are present together in a multi-dose container. The composition may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles such as aqueous polyethylene glycol solutions. Examples of oily or non-aqueous vehicles, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate), and may contain, for example, preservatives, A formulation of wetting agents, emulsifiers or suspending agents, stabilizers and/or dispersing agents. Alternatively, the active ingredient may be in the form of a powder obtained by sterile separation of sterile solids or by lyophilization from solution for reconstitution with a suitable vehicle (for example, sterile pyrogen-free water) before use.

The compounds of the invention may be formulated as ointments, creams or lotions, or as transdermal patches for topical administration to the epidermis. Ointments and creams can be formulated, for example, with an aqueous or oily base with the addition of a suitable thickening and/or gelling agent. The lotion may be formulated with an aqueous or oily base and will usually also contain one or more emulsifiers, stabilizers, dispersing agents, suspending agents, thickening agents or coloring agents. Formulations suitable for topical administration in the mouth include: buccal agents comprising the active agent in a flavoring base (usually sucrose and gum arabic or tragacanth); tablets comprising an inert matrix (such as gelatin and glycerin) Or an active ingredient in sucrose and gum arabic; and a mouthwash comprising the active ingredient in a suitable liquid carrier.

The compounds of the invention may be formulated as a suppository for administration. The low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted and the active component is uniformly dispersed, for example, by stirring. The molten homogeneous mixture is then poured into a mold of suitable size which is allowed to cool and solidify.

The compounds of the invention may be formulated for vaginal administration. Pests, tampons, creams, gels, pastes, foams or sprays containing such carriers as known in the art, in addition to the active ingredient, are suitable.

The compounds of the invention may be formulated for nasal administration. By conventional means (for example, A dropper, pipette or sprayer) applies the solution or suspension directly to the nasal cavity. Formulations may be provided in single or multiple doses. In the latter condition after the dropper or straw, this can be accomplished by administering to the patient a suitable predetermined volume of solution or suspension. In the case of a nebulizer, this can be achieved, for example, by means of a metered atomizing jet pump.

The compounds of the present invention may be formulated as an aerosol for administration to the respiratory tract, and the aerosol administration includes intranasal administration. The compound will typically have a small particle size of, for example, about 5 microns or less. The particle size can be obtained by means known in the art, for example by micronization. The active ingredient is provided in a pressurized pack with a suitable propellant such as chlorofluorocarbon (CFC) (eg dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane) or carbon dioxide or Other suitable gases. Aerosols also preferably contain a surfactant such as lecithin. The drug dose can be controlled by a metering valve. Alternatively, the active ingredient may be presented in the form of a dry powder, such as a powder mixture in a suitable powder base such as lactose, starch, starch derivatives (such as hydroxypropyl methylcellulose) and polyvinylpyrrolidone ( PVP). The powder carrier will form a gel in the nasal cavity. The powder composition may be presented in unit dosage form in a capsule or cartridge such as a gelatin or blister pack, wherein the powder may be administered by an inhaler.

Formulations having an enteric coating suitable for sustained or controlled release administration of the active ingredient can be prepared, if desired. For example, the compounds of the invention can be formulated in a transdermal or subcutaneous drug delivery device. Such delivery systems are advantageous when sustained release of the compound is necessary and when patient compliance with the treatment regimen is critical. Compounds in transdermal delivery systems are typically attached to a skin-adhesive solid support. The relevant compound can also be combined with a penetration enhancer such as azone (1-dodecylaza-cycloheptan-2-one). The sustained release delivery system is inserted subcutaneously into the subcutaneous layer by surgery or injection. The subcutaneous implant encapsulates the compound in a lipid soluble film, such as a polyoxyxene rubber, or a biodegradable polymer, such as polyacetic acid.

Suitable formulations as well as pharmaceutical carriers, diluents and excipients are described in Remington: The Science and Practice of Pharmacy 1995, edited by EW Martin, Mack Publishing Company, 19th edition, Easton, Pennsylvania. A skilled blending scientist can modify the formulation under the teachings of the specification to provide a variety of formulations for a particular route of administration without causing instability or compromised therapeutic activity of the compositions of the present invention.

In order to make the compounds of the present invention more soluble in water or other vehicles, the changes to such compounds can be readily accomplished, for example, by minor modifications (salt formulations, esterification, etc.), which are fully within the skill of the art. Within the scope. It is also within the general skill of the art to modify the route of administration of a particular compound and the dosing regimen to control the pharmacokinetics of the compounds of the invention to produce maximum beneficial effects in the patient.

The term "therapeutically effective amount" as used herein means the amount required to alleviate the symptoms of a disease in an individual. The dosage can be adjusted as needed in each particular situation. Depending on the severity of the disease to be treated, the age and general health of the patient, the other drug being treated, the route and form of administration, and the various factors of the practitioner's preferences and experience, the dose may vary within wide limits. . For oral administration, in monotherapy and/or combination therapy, a daily dose of between about 0.01 and about 1,000 mg per kilogram of body weight per day should be appropriate. The daily dose is preferably between about 0.1 and about 500 mg per kilogram of body weight per day, more preferably between 0.1 and about 100 mg per kilogram of body weight per day, and optimally at 1.0 and about 10 mg per kilogram of body weight per day. between. Thus, for administration to a 70 kg individual, the dosage range can be from about 7 mg to 0.7 g per day. The daily dose can be administered in a single dose or in divided doses, usually between 1 and 5 doses per day. Generally, treatment is initiated with a smaller dose that is less than the optimal dose of the compound. Subsequently, the dose is increased in small increments until the individual patient achieves the best results. Those of ordinary skill in the art will be able to determine the therapeutically effective amount of a compound of the present invention for a given disease and patient, without undue experimentation, relying on personal knowledge, experience, and disclosure of the present application.

The pharmaceutical preparation is preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation, the package containing discrete amounts Formulations, such as packaged tablets, capsules and vials or ampoules. Further, the unit dosage form can be a capsule, a lozenge, a troche, or a buccal preparation itself, or it can be a suitable number of such unit dosage forms in the form of a package.

Indications and treatments

The compound of formula I inhibits Bruton's tyrosine kinase (Btk). Phospholipase-Cy is activated by activation of Btk by an upstream kinase, which in turn stimulates the release of pro-inflammatory mediators. Compounds of formula I incorporating a 1-sided oxy-1H-pyridazin-2-yl side chain exhibit unexpectedly enhanced inhibitory activity compared to analogs having other side chains. The hydroxymethyl substitution on the phenyl ring provides unexpectedly enhanced efficacy compared to analogs having an alternative substitution at this position. The compounds of formula I are useful in the treatment of arthritis and other anti-inflammatory and autoimmune diseases. The compounds of formula I are therefore suitable for the treatment of arthritis. The compounds of formula I are useful for inhibiting Btk in cells and for modulating B cell development. The invention further comprises a pharmaceutical composition comprising a mixture of a compound of formula I and a pharmaceutically acceptable carrier, excipient or diluent.

The compounds described herein are kinase inhibitors, in particular, Btk inhibitors. Such inhibitors are useful in the treatment of one or more diseases which are responsive to kinase inhibition, including diseases which inhibit Btk inhibition in mammals and/or inhibit B cell proliferation. Without wishing to be bound by any particular theory, it is believed that the interaction of a compound of the invention with Btk inhibits Btk activity and thus produces the pharmaceutical utility of such compounds. Accordingly, the invention encompasses treating a mammal (eg, a human) having a disease that inhibits Btk activity and/or inhibits B cell proliferation. A method comprising administering to a mammal having the disease an effective amount of at least one of the chemical entities provided herein. The effective concentration can be determined experimentally, for example, by analyzing the blood concentration of the compound or theoretically by calculating the bioavailability. Other kinases that may be affected in addition to Btk include, but are not limited to, other tyrosine kinases and serine/threonine kinases.

Kinases play a significant role in signaling pathways that control essential cellular processes such as proliferation, differentiation, and death (apoptosis). Abnormal kinase activity has been implicated in a variety of diseases, including a variety of cancers, autoimmune and/or inflammatory diseases, and acute inflammatory responses. The role of kinases in key cellular signaling pathways provides an important opportunity to identify novel drugs that target kinases and signaling pathways.

One embodiment includes a method of treating a patient having an autoimmune and/or inflammatory disease, or having an acute inflammatory response to inhibition of Btk activity and/or B cell proliferation.

Autoimmune and/or inflammatory diseases that may be affected by the use of the compounds and compositions of the invention include, but are not limited to, psoriasis, allergies, Crohn's disease, colonic urgency, and Hugh's disease. (Sjogren's disease), tissue transplant rejection and acute rejection of transplanted organs, asthma, systemic lupus erythematosus (and related spheroid nephritis), dermatomyositis, multiple sclerosis, scleroderma, vasculitis ( ANCA-related and other vasculitis), autoimmune hemolytic and thrombocytopenic states, Goodpasture's syndrome (and related spheroid nephritis and pulmonary hemorrhage), atherosclerosis, rheumatoid arthritis, Chronic idiopathic thrombocytopenic purpura (ITP), Addison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock, and myasthenia Powerless.

Included herein are methods of treatment wherein at least one of the chemical entities provided herein is administered in combination with an anti-inflammatory agent. Anti-inflammatory agents include, but are not limited to, NSAIDs, non-specific and COX-2 specific epoxidase inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor receptor (TNF) receptors Antagonists, immunosuppressants and Methamidine.

Examples of NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, diclofenac sodium and misoprostol (misoprostol) a combination of sulindac, oxaprozin, diflunisal, piroxicam, indomethacin, etodolac, Fenoprofen calcium, ketoprofen, sodium nabumetone, sulfasalazine, tolmetin sodium, and hydroxychloroquine. Examples of NSAIDs also include COX-2 specific inhibitors, such as celecoxib, valdecoxib, lumiracoxib, and/or etoricoxib.

In some embodiments, the anti-inflammatory agent is a salicylate. Salicylates include, but are not limited to, acetylsalicylic acid or aspirin, sodium salicylate, and choline and magnesium salicylate.

The anti-inflammatory agent can also be a corticosteroid. For example, the corticosteroid can be corticosterone, dexamethasone, methylprednisolone, prednisolone, sodium nylon phosphate or prednisone.

In other embodiments, the anti-inflammatory agent is a gold compound such as sodium gold thiomalate or auranofin.

The invention also includes embodiments in which the anti-inflammatory agent is a metabolic inhibitor, such as a dihydrofolate reductase inhibitor, such as a methotrexate or a dihydroorotate dehydrogenase inhibitor, such as leflunomide. .

Other embodiments of the invention pertain to combinations wherein at least one anti-inflammatory compound is an anti-C5 monoclonal antibody (such as eculizumab or pexelizumab), a TNF antagonist (such as etasie) Enantercept or infliximab (which is an anti-TNFα monoclonal antibody).

Other embodiments of the invention relate to combinations wherein at least one active agent is immunized An inhibitory compound, such as an immunosuppressive compound selected from the group consisting of methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine, and mycophenolate morpholine B Ester (mycophenolate mofetil).

B cells and B cell precursors that express Btk have been associated with the pathology of B cell malignancies including, but not limited to, B cell lymphomas, lymphomas (including Hodgkin's lymphoma). And non-Hodgkin's lymphoma, hairy cell lymphoma, multiple myeloma, chronic and acute myeloid leukemia, and chronic and acute lymphocytic leukemia.

BTK has been shown to be an inhibitor of Fas/APO-1 (CD-95) death-inducing signaling complex (DISC) in B-lineage lymphoid cells. The end result of leukemia/lymphoma cells may be present in the balance between the anti-apoptotic effect of DISC-activated caspase and the upstream anti-apoptotic regulatory mechanisms involved in BTK and/or its receptor (Vassilev) Et al., J. Biol . Chem . 1998, 274, 1646-1656).

BTK inhibitors have also been found to be useful as chemosensitizers and are therefore suitable for use in combination with other chemotherapeutic agents, particularly those that induce apoptosis. Examples of other chemotherapeutic agents that can be used in combination with chemically sensitizing BTK inhibitors include type I topoisomerase inhibitors (camptothecin or topotecan), type II topoisomerase inhibitors (eg, Daunomycin and etoposide), alkylating agents (eg cyclophosphamide, melphalan and BCNU), tubulin targeting agents (eg taxol and vinblastine) (vinblastine)), and biological agents (eg, antibodies such as anti-CD20 antibodies, IDEC 8, immunotoxins, and cytokines).

Btk activity is also associated with some leukemias that express a bcr-abl fusion gene caused by a partial translocation of chromosomes 9 and 22. This anomaly is usually observed in chronic myelogenous leukemia. Btk is constitutively phosphorylated by bcr-abl kinase, which initiates downstream survival signals, thereby preventing apoptosis in bcr-abl cells (N. Feldhahn et al., J. Exp. Med. 2005 201(11): 1837-1852 ).

treatment method

The application provides a method of treating an inflammatory and/or autoimmune condition comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a method of treating an inflammatory condition comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a method of treating rheumatoid arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The application provides a method of treating an inflammatory and/or autoimmune condition comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I.

The application provides a method of treating arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I.

The application provides a method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I.

The application provides a method of inhibiting B cell proliferation comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I.

The present application provides a method of inhibiting Btk activity comprising administering a Btk inhibitor compound of any of Formula I, wherein the Btk inhibitor compound exhibits a concentration of 50 micromolar or 50 micromolar in an in vitro biochemical assay of Btk activity. IC ₅₀ below the ear concentration.

In a variant of the above method, Btk inhibitor compound exhibits a concentration of 100 or 100 nemorubicin nemorubicin ear ear below the concentration of IC ₅₀ in an in vitro biochemical assay of Btk activity in the.

In another variant of the above method, the compound exhibits a concentration of 10 or 10 nemorubicin nemorubicin ear ear below the concentration of IC ₅₀ in an in vitro biochemical assay of Btk activity in the.

The application provides a method of treating an inflammatory condition comprising co-administering to a patient in need thereof a combination of a therapeutically effective amount of an anti-inflammatory compound with a Btk inhibitor compound of Formula I.

The application provides a method of treating arthritis comprising co-administering to a patient in need thereof a combination of a therapeutically effective amount of an anti-inflammatory compound with a Btk inhibitor compound of Formula I.

The present application provides a method of treating lymphoma or BCR-ABL1 ⁺ leukemia cells by administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of Formula I.

Instance abbreviation

Commonly used abbreviations include: acetonitrile (Ac), azo-bis-isobutyl decyl nitrile (AIBN), atmospheric pressure (Atm), 9-boron bicyclo [3.3.1] decane (9-BBN or BBN), 2, 2 '-Bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), tert-butoxycarbonyl (Boc), di-tert-butyl dicarbonate or boc anhydride (BOC ₂ O), benzene Base (Bn), butyl (Bu), Chemical Abstracts Registration Number (CASRN), benzyloxycarbonyl (CBZ or Z), carbonyl diimidazole (CDI), 1,4-diazabicyclo [2.2.2] Octane (DABCO), diethylaminosulfur trifluoride (DAST), dibenzylideneacetone (dba), 1,5-diazabicyclo[4.3.0]壬-5 -ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), N,N'-dicyclohexylcarbodiimide (DCC), 1,2-di Ethyl chloride (DCE), dichloromethane (DCM), 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), diethyl azodicarboxylate (DEAD), Di-isopropyl azodicarboxylate (DIAD), diisobutylaluminum hydride (DIBAL or DIBAL-H), di-isopropylethylamine (DIPEA), N,N-dimethylacetamide (DMA) ), 4-N,N-dimethylaminopyridine (DMAP), N,N-dimethylformamide (DMF), dimethyl hydrazine (DMSO), 1,1'-bis-(diphenyl Ethyl (dppe), 1,1'-bis-(diphenylphosphino)ferrocene (dppf), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride (EDCI), 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), ethyl (Et), ethyl acetate (EtOAc), ethanol (EtOH), Ethyl 2-ethoxy-2H-quinoline-1-carboxylate (EEDQ), diethyl ether (Et ₂ O), isopropoxy ethane (EtOiPr), hexafluorophosphate O-(7-azabenzotriene) Zin-1-yl)-N,N,N',N'-four (HATU), acetic acid (HOAc), 1-N-hydroxybenzotriazole (HOBt), high pressure liquid chromatography (HPLC), isopropanol (IPA), isopropylmagnesium chloride (iPrMgCl), hexamethyldi Hydrazine alkane (HMDS), liquid chromatography mass spectrometry (LCMS), lithium hexamethyldiazepine (LiHMDS), m-chloroperoxybenzoic acid (m-CPBA), methanol (MeOH), melting point (mp ), MeSO ₂ -(methylsulfonyl or Ms), methyl (Me), acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA), mass spectrometry (ms), methyl tert-butyl ether (MTBE), Methyltetrahydrofuran (MeTHF), N-bromosuccinimide (NBS), n-butyllithium (nBuLi), N-carboxy anhydride (NCA), N-chlorobutaneimine (NCS), N-A Phenylmorpholine (NMM), N-methylpyrrolidone (NMP), pyridinium chlorochromate (PCC), dichloro-((bis-diphenylphosphino)ferrocenyl)palladium(II) (Pd (dppf)Cl ₂ ), palladium(II) acetate (Pd(OAc) ₂ ), ginseng (diphenylmethyleneacetone) dipalladium (0) (Pd ₂ (dba) ₃ ), dichromate pyridinium (PDC) ), phenyl (Ph), propyl (Pr), isopropyl (i-Pr), pounds per square foot (psi), pyridine (pyr), 1,2,3,4,5-pentaphenyl- 1'-(di-tert-butylphosphino)ferrocene (Q-Phos), room temperature (ambient temperature, rt or RT), second butyl lithium (sBuLi), third Alkyl dimethyl silicone or _{t-BuMe 2 Si (TBDMS)} , tetra - n-butylammonium (TBAF), triethylamine (TEA or Et ₃ N), 2,2,6,6- tetramethyl- Piperidine 1-alkoxy (TEMPO), trimethyldecyl ethoxymethyl (SEM), triflate or CF ₃ SO ₂ -(Tf), trifluoroacetic acid (TFA), 1 , 1'-bis-2,2,6,6-tetramethylheptane-2,6-dione (TMHD), tetrafluoroborate O-benzotriazol-1-yl-N,N,N' , N'-four (TBTU), thin layer chromatography (TLC), tetrahydrofuran (THF), trimethyldecyl or Me ₃ Si (TMS), p-toluenesulfonic acid monohydrate (TsOH or pTsOH), 4-Me-C ₆ H ₄ SO ₂ - or tolsulfonyl (Ts) and N-urethane-N-carboxy anhydride (UNCA). Conventional nomenclatures including prefixes (n), iso (i-), second (sec-), third (tert-), and neo (neo) have their usual use when used with alkyl moieties. Meaning (J. Rigaudy and DP Klesney, Nomenclature in Organic Chemistry, IUPAC 1979 Pergamon Press, Oxford.).

General condition

The compounds of the present invention can be prepared starting from commercially available starting materials using conventional synthetic techniques and procedures known to those skilled in the art. The following summary is applicable to the preparation of such compounds. Reaction process. Other illustrative illustrations can be found in a particular example.

Preparation example

6-t-butyl-2H-phthalazin-1-one

In 1 L round bottom flask, 4-tert-butyl benzoyl chloride (97.2 g, 90 ml, 494 mmol, eq: 1.00) and CH ₂ Cl ₂ (375 ml) were combined to give a colorless solution. Cool to 0 °C. 2-Methylpropan-2-amine (79.5 g, 114 ml, 1.09 mol, equivalent: 2.2) was added dropwise from a dropping funnel, keeping the temperature below 10 °C. Rinse the side of the addition funnel with more CH ₂ Cl ₂ . The reaction was warmed to 25 °C over 3 hours. 4 N NaOH was added slowly until the solids dissolved (about 300 mL). Separate the layers. The aqueous layer was extracted with 4× CH ₂ Cl ₂ . The organic layer (total 2.5 L) dried over Na ₂ SO ₄ and concentrated to about 250 mL. A large amount of precipitate is formed. The flask was cooled in an ice bath. The resulting white solid was filtered and the white solid obtained was needle (72 g) was washed and 3X CH ₂ Cl ₂ 1X EtOAc, dried under vacuum pumped dried. The combined filtrate and washings were again concentrated to 100 mL. More crystals are formed. cool down. Second crop was filtered and the resulting white solid (31.6 g) was washed with ₂ Cl ₂ CH. ¹ H NMR (300 MHz, chloroform - d ) δ ppm 1.33 (s, 9 H) 1.47 (s, 9 H) 5.91 (br.s., 1 H) 7.43 (d, J = 8.31 Hz, 2 H) 7.66 (d, J = 8.31 Hz, 2 H)

This reaction was carried out in batches of 2 x 26 g and combined. For each batch, a 2 L three-necked flask equipped with a mechanical stirrer, an addition funnel connected to the bubbler via a tube, and a nitrogen inlet was used. The system was heated in an oven and cooled under a stream of N _2. N,4-Di-t-butylbenzamide (26.2 g, 112 mmol, eq. 1.00) was combined with THF (1.01 1) to give a colourless solution. The reaction mixture was cooled to -78 °C. The cyclohexane containing the second butyllithium (176 ml, 1.4 M, 247 mmol, equivalent: 2.2) was slowly added dropwise under a stream of N ₂ . A yellow solution was obtained. The reaction mixture was warmed to -15 °C over 1 hour. A yellow suspension was obtained. The reaction mixture was cooled back to -78 °C. Anhydrous DMF (16.4 g, 17.4 ml, 225 mmol, eq. 2) was added dropwise. The reaction was warmed to 0 °C. LCMS indicated the reaction was complete.

150 mL of saturated NH ₄ Cl was slowly added at 0 °C. The reaction mixture was allowed to stand at ambient temperature and then transferred to a 4 L conical flask and rinsed with water and EtOAc. The two batches of the reaction were combined and then transferred to the same 4 L Erlenmeyer flask.

The total volume of the two batches of the reaction mixture was slightly more than 3 L. The reaction mixture was portioned in a 2 L round-bottomed flask, and concentrated on a rotary evaporator at 50 ° C to avoid boiling. The volume is reduced to approximately 200-300 mL.

There is a large amount of solid precipitate. LCMS indicated the absence of product in the liquid portion. A white solid was collected by filtration in a large sintered glass funnel. The chunks were suspended in water in a funnel and chopped into fine powder with a crowbar and filtered. The solid was washed 5 times with water and then dried in a vacuum oven at 50 ° C for 2 days to give 59.6 g of 2,5-di-t-butyl-3-hydroxyisoindolin-1-one as a white solid. ¹ H NMR was carried out and used as it is in the next step. ¹ H NMR (300 MHz, chloroform - d ) δ ppm 7.27 (s, 9 H) 7.53 (s, 9 H) 8.26 (d, J = 11.71 Hz, 1 H) 11.90 (d, J = 11.71 Hz, 1 H 13.40-13.46 (m, 1 H) 13.47 (s, 1 H) 13.51-13.58 (m, 1 H).

2,5-di-t-butyl-3-hydroxyisoindol-1-one (59.6 g, 228 mmol, in a 2 L three-necked flask equipped with an addition funnel, air cooler and nitrogen inlet) Equivalent: 1.00) combined with acetic acid (868 ml) gave a pale yellow solution. The reaction was heated to 90 °C. Monohydrate monohydrate (14.8 g, 14.4 ml, 296 mmol, equivalent: 1.3) was added dropwise. The reaction mixture was stirred at 90 ° C for 1 hour, then the reaction mixture was diluted with 300 mL H ₂ O and slowly cooled to 25 ° C. LCMS confirmed the completion of the reaction and was completely converted to the desired product. The reaction was concentrated to a reduced volume (about 50-100 mL). A colorless solid was produced in the solution after standing. The solid was collected by EtOAc (EtOAc)EtOAc. . ¹ H NMR (400 MHz, chloroform - d ) δ ppm 1.44 (s, 9 H) 7.66-7.73 (m, 1 H) 7.84 - 7.91 (m, 1 H) 8.18 (d, J = 0.51 Hz, 1 H) 8.33-8.42 (m, 1 H).

2-(6-Tert-butyl-1-oxo-1H-pyridazin-2-yl)-6-chloro-benzaldehyde

In a 500 mL three-necked flask equipped with a N ₂ inlet and a bubbler outlet and a mechanical stirrer, 6-t-butylpyridazine-1(2H)-one (6.46 g, 31.9 mmol, eq: 1.00) 2-Chloro-6-fluorobenzaldehyde (7.6 g, 47.9 mmol, equivalent: 1.5) and potassium carbonate (8.83 g, 63.9 mmol, equivalent: 2) (fine powder) combined with DMA (70 ml) to give a yellow suspension . Tetraethylammonium chloride (688 mg, 4.15 mmol, equivalent: 0.13) was added and the reaction mixture was warmed at 68 ° C (bath temperature) with vigorous stirring. After the reaction mixture became a solution, it was stirred at this temperature for 1 hour. The reaction was continued to warm overnight at 75 ° C (bath temperature). Further, 2-chloro-6-fluorobenzaldehyde (5 g, 31.9 mmol, equivalent: 1.00) was added and the reaction mixture was warmed at 75 ° C (bath temperature) for 3 days. The reaction mixture was poured and the (5 × 300 mL) and extracted with CH ₂ Cl ₂ into 300 mL H ₂ O in. The organic layer was dried over Na ₂ SO ₄ and concentrated in vacuo. A large amount of DMA remains. The reaction mixture was poured into 500 mL H ₂ O and extracted with ether (5 × 300 mL). , The ether layer was washed with brine and dried over MgSO _4, filtered and concentrated in vacuo. When the volume is about 150 mL, a large amount of white crystalline material is present. The product was isolated by filtration to give the pure product (5.4 g). The filtrate was concentrated in vacuo and purified using an Analogix system by SF25-160 g column (with 5% -25% (3: 1 CH 2 Cl 2 / EtOAc) / hexanes eluting) the residue was purified to give additional 4.75 g of desired product. The total yield of 2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6-chloro-benzaldehyde was 10.1 g (93%). ¹ H NMR (300 MHz, chloroform - d ) δ ppm 7.27 (s, 9 H) 13.30 (d, J = 7.55 Hz, 1 H) 13.34-13.51 (m, 2 H) 13.56 (d, J = 1.51 Hz, 1 H) 13.71 (dd, J = 8.50, 1.70 Hz, 1 H) 14.12 (s, 1 H) 14.21 (d, J = 8.31 Hz, 1 H) 16.20 (s, 1 H). LC / MS observations ^{[M + H] + 340.8;} [M + Na] + 362.9.

6-t-butyl-2-(3-chloro-2-hydroxymethyl-phenyl)-2H-phthalazin-1-one

To 2-(6-t-butyl-1-oxo-oxazine-2(1H)-yl)-6-chlorobenzaldehyde (61 g, at 0 ° C under a nitrogen stream with vigorous stirring over 20 min. 179 mmol, eq.: 1.00) 1 g of sodium borohydride (7.45 g, 197 mmol, eq.: 1.1) was added in portions in CH ₂ Cl ₂ (350 ml) and methanol (350 ml). The reaction mixture was warmed to ambient temperature and stirred at this temperature for 1 hour. Water (100 mL) was added dropwise, followed by CH ₂ Cl ₂ and more water and the layers were separated. The aqueous phase was extracted 3 times with CH ₂ Cl ₂ . Combined organic layers were washed with saturated NH ₄ Cl and dried over sodium sulfate. After filtration, the yellow liquid was concentrated to give a yellow-green solid. This solid was triturated with diethyl ether / ethyl acetate (6:1) and washed several times with the same solvent to afford a pale yellow-green solid. This solid was slurried in dichloromethane containing 1% methanol. The resulting white solid was collected by EtOAc (EtOAc m.) -2H-phthalazin-1-one (28.3 g). ¹ H NMR (300 MHz, chloroform - d ) δ ppm 1.45 (s, 9 H) 4.58 (br.s., 2 H) 7.31 (dd, J = 7.93, 1.51 Hz, 1 H) 7.43 (t, J = 7.93 Hz, 1 H) 7.57 (dd, J == 8.12, 1.32 Hz, 1 H) 7.77 (d, J = 1.89 Hz, 1 H) 7.93 (dd, J = 8.69, 1.89 Hz, 1 H) 8.33 (s , 1 H) 8.44 (d, J = 8.69 Hz, 1 H); LC/MS observed [M+H] ⁺ 343.0.

2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6-chloro-phenylmethyl acetate

To 6-tert-butyl-2-(3-chloro-2-(hydroxymethyl)phenyl)pyridazine-1(2H)-one (28.3 g, 82.6 mmol, eq: 1.00) in dichloromethane ( Triethylamine (10.9 g, 10.1 ml, 107 mmol, equivalent: 1.3), acetic anhydride (11.0 g, 15.1 ml, 107 mmol, equivalent: 1.3) and DMAP (252 mg, 2.06 mmol) were added to the solution in 250 ml). , equivalent: 0.025) and the solution was stirred overnight at ambient temperature. The reaction mixture was poured into 300 mL H ₂ O and extracted with _{CH 2 Cl 2 (3 × 250} mL).

The organic layer was dried over sodium sulfate, filtered and evaporatedEtOAc The semisolid was dissolved in 300 mL of isopropyl acetate and concentrated to give a pale yellow solid. This solid was wet-milled with 150 mL of isopropyl acetate under cooling, then filtered and washed once with 30 mL of cold isopropyl acetate and then washed with 50 mL of 2:1 hexanes: isopropyl acetate to afford white Fluorously solid 2-(6-tert-butyl-1-oxooxy-1H-pyridazin-2-yl)-6-chloro-benzyl ester (23.9 g). ¹ H NMR (300 MHz, chloroform - d ) δ ppm 1.44 (s, 9 H) 1.91 (s, 3 H) 5.18 (s, 2 H) 7.31 - 7.38 (m, 1 H) 7.45 (t, J = 7.93) Hz, 1 H) 7.51 - 7.59 (m, 1 H) 7.73 (d, J = 1.89 Hz, 1 H) 7.89 (dd, J = 8.69, 1.89 Hz, 1 H) 8.25 (s, 1 H) 8.40 (d) , J = 8.69 Hz, 1 H). LC/MS observed [M+H] ⁺ 385.0.

2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1,3,2]diacetate Oxyboron -2-yl)-benzyl ester

To a 1 L round bottom flask was added 2-(6-t-butyl-1-oxooxazin-2(1H)-yl)-6-chlorobenzyl ester (16.4 g, 42.6 mmol, eq. 1.00) and bis-pinacol borate (20.6 g, 81.0 mmol, equivalent: 1.9). The material was dissolved in methyltetrahydrofuran (300 ml) and stirred for 5 min. The mixture was evacuated and backfilled three times with argon, then potassium acetate (10.5 g, 107 mmol, eq. Palladium(II) acetate (239 mg, 1.07 mmol, equivalent: 0.025) was added. X-PHOS (1.02 g, 2.13 mmol, eq. 0.05) was added. The mixture was evacuated and backfilled three times with argon. The orange-yellow mixture was heated to 68 ° C (bath temperature) for 1 hour. Only a small amount of the desired product was formed after 1 hour. The reaction temperature was adjusted to 70 °C. The progress of the reaction was closely monitored by LCMS. The reaction was stirred at this temperature for a further 9 hours, then only a small amount of starting material, traces of &lt The reaction was stirred at 70 ° C for an additional hour to promote completion of the reaction. The color turns brown. The reaction was cooled to ambient temperature. And the reaction mixture was poured (3 × 250 mL) and extracted with EtOAc into 400 mL H ₂ O in. The combined organic extracts were washed with brine, dried (MgSO _4), filtered and concentrated in vacuo. The residue was purified using an Analogic purification system eluting with a SF 25-600 g column eluting with 100% hexane for 15 min then eluting with 0-25% EtOAc/hexanes. The intermediate fraction of the peak of the product was concentrated, and 18 g of 2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6 was obtained as a yellow foam. -(4,4,5,5-tetramethyl-[1,3,2]diboron 2-yl)-benzyl ester (purity >95%). ¹ H NMR (300 MHz, chloroform - d ) δ ppm 1.34 (s, 12 H) 1.44 (s, 9 H) 1.87 (s, 3 H) 5.30 (br.s., 2 H) 7.39-7.57 (m, 2 H) 7.72 (d, J = 1.51 Hz, 1 H) 7.78 (dd, J = 8.50, 1.70 Hz, 1 H) 7.97 (dd, J = 6.42, 2.64 Hz, 1 H) 8.24 (s, 1 H) 8.40 (d, J = 8.31 Hz, 1 H). LC/MS observations for the main LC peaks [M+H] ⁺ 477. It should be noted that this material is prepared several times and the purity of the desired product is in the range of from about 50% to about 95%.

6-fluoro-2H-phthalazin-1-one

N,N-Diethyl-4-fluoro-2-methylindolyl-benzamide (2.94 g, 13.2 mmol), hydrazine monohydrate (Sigma-Aldrich; 725 mg, 14.5 mmol) and EtOH (15 mL) The mixture was heated in a large capacity microwave tube at 50 ° C for 45 minutes. Ice AcOH (2 mL) was added and the mixture was heated in a Biotage Initiator microwave at 150 °C for 2 hours. Was added _saturated aqueous NaHCO (75 mL) and EtOAc (70 mL). The organic layer was washed successively with a 5% aqueous NaHCO ₃ solution and a 50% brine solution. The aqueous layer was back extracted with EtOAc (2×60 mL). Dried (MgSO ₄₎ the organic layers were combined, filtered and evaporated to give a pale yellow powder of 6-fluoro--2H- phthalazin-1-one (2.14 g, 99%). LC/MS observed [M+H] ⁺ 165.

2-methyl-2-(1-o-oxy-1,2-dihydro-pyridazin-6-yl)-propanenitrile

Potassium bis(trimethyldecyl)amine (0.91 M in THF, Alfa-Aesar; 46.6 mL, 42.4 mmol) was added to 6-fluoro-2H-phthalazin-1-one (1.16 g, 7.1 mmol) In a solution of THF (20 mL). The mixture was heated overnight at 70 ° C in an oil bath. Water (125 mL) and EtOAc (100 mL) were added. The layers were separated and the aqueous extracted with EtOAc EtOAc Dried (MgSO ₄₎ the organic layers were combined, filtered and evaporated to give a yellow solid (1.52 g). By CH _2/2 Cl triturated with hot hexane, followed by filtration to purify this material to give a pale yellow powder of 2-methyl-2- (1-oxo-1,2-dihydro - phthalazin Pyrazin-6-yl)-propionitrile (1.12 g, 74%). LC/MS observed [M+H] ⁺ 214.

2-[2-(3-Bromo-2-methylindenyl-phenyl)-1-oxooxy-1,2-dihydro-pyridazin-6-yl]-2-methyl-propanenitrile

2-Methyl-2-(1-o-oxy-1,2-dihydro-pyridazin-6-yl)-propanenitrile (1.12 g, 5.23 mmol), 2,6-dibromobenzaldehyde (2.21) _{g, 8.37 mmol), NaHCO 3} (879 mg, 10.5 mmol), a mixture of copper (I) iodide (996 mg, 5.23 mmol) and DMSO (35.3 mL) was heated the at about 110 ℃ 2 hours. The mixture was cooled to room temperature. CH ₂ Cl ₂ (40 mL) and water (40 mL) were added. The mixture was filtered through celite and the celite was washed thoroughly with CH ₂ Cl ₂ and MeOH. The layer in the filtrate was separated and the organic layer was washed with 50% brine. The aqueous layer was back-extracted with _{CH 2 Cl 2 (2 × 40} mL). Combined organic layers were dried (MgSO _4), filtered and concentrated in vacuo. Purification by Analogix system using silicon dioxide SF25-60 g column (with 0-5% MeOH / CH ₂ Cl ₂ fractions) to give the residue, to give a pale yellow solid of 2- [2- (3-bromo - 2-Methylmethyl-phenyl)-1-oxo-1,2-dihydro-pyridazin-6-yl]-2-methyl-propanenitrile (1.89 mg, 82%), purity about 90 %. Observed for LC/MS [M+H] ⁺ 396/398.

Example 1

2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl -6-Phenoxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-6-tert-butyl-2H-phthalazin-1-one

2-(6-T-butyl-1-oxo-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1, 3,2]diboron -2-yl)-benzyl ester (intermediate 5; 177 mg, 371 μmol), 4-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy )-pyridin-2-ylamino]-6-chloro-2-methyl-2H-pyridazin-3-one (which can be as described in Step 2 of Berthel, SJ et al., US 20120040949, I-13) Description Preparation; 90 mg, 247 μmol), 2-dicyclohexylphosphine-2', 4',6'-triisopropylbiphenyl (18 mg, 37 μmol), tripotassium phosphate (144 mg, 0.68 mmol) A mixture of water (332 μL) and n-butanol (1.4 mL) was degassed and backfilled with argon.

Bis(dibenzylideneacetone)palladium(0) (10 mg, 17 μmol) was added and the mixture was degassed as previously described. The mixture was heated at 110 ° C for 2.5 hours. Water (35 mL) and EtOAc (35 mL) were added and mixture was shaken in a sep. funnel. The organic layer was collected and washed with brine (35 mL). The aqueous layer was back extracted with EtOAc (2×30 mL). The organic layer was dried (MgSO ₄₎ the combined, filtered, evaporated and purified using an Analogix system (with _{1% -20% MeOH / CH 2} Cl 2 fractions) was purified by column 24g to give a viscous pale yellow oil of acetic acid 2-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl-6- Sideoxy-1,6-dihydro-pyridazin-3-yl}-6-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-phenylmethyl ester with 2 -(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl- Mixture of 6-o-oxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-6-tert-butyl-2H-phthalazin-1-one (112 Mg). Dioxane (572 μL) and 2 N aqueous NaOH (980 μL, 1.96 mmol) were added and the mixture was heated at 50 ° C for 2 hours under argon. Water (25 mL) and EtOAc (50 mL) were added and the mixture was shaken. The organic layer was collected and washed with brine (50 mL). The aqueous layer was back extracted with EtOAc (2×40 mL). Dried (MgSO ₄₎ combined organic layers were, filtered, evaporated and purified by column 12g (with _{1% -15% MeOH / CH 2} Cl 2 fractions) using Analogix purification system, to give a pale yellow solid of 2- ( 3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl-6- Sideoxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-6-tert-butyl-2H-phthalazin-1-one (81 mg, 52% ). LC/MS observed [M+H] ⁺ 636.0.

Example 2

6-Tertibutyl-2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6' -hexahydro- [3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-2H-pyridazin-1-one

Purging 2-(6-t-butyl-1-oxooxy-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1, 3,2]diboron 2-yl)-benzyl ester (which can be prepared as described in Example 5; about 75% purity; 214 mg, about 0.34 mmol), 6-chloro-2-methyl-4-(5-(1- Methylpiperidin-4-yl)pyridin-2-ylamino)pyridazine-3(2H)-one (which can be prepared as described in Berthel, SJ et al, US 20120040949, Example 56; 100 mg; , 0.3 mmol), bis(dibenzylideneacetone)palladium (Aldrich; 8.6 mg, 15 μmol), XPhos (2-(dicyclohexylphosphino)-2', 4',6'-triisopropyl -1,1'-biphenyl, Strem Chemicals; 14.3 mg, 30 μmol) and a mixture of tripotassium phosphate (159 mg, 0.75 mmol) with degassed n-butanol (2.4 mL) and water (600 μL) mixture. The mixture was heated overnight at 115 ° C in a sand bath. Diatomaceous earth mixture was filtered, and concentrated by chromatography (eluted with 0-25% MeOH / CH ₂ Cl ₂ fractions) to give a yellow oil (150 mg). MeOH (2.3 mL) and potassium carbonate (96 mg, 0.7 mmol) were added and the mixture was stirred at 45 ° C for one hour. CH ₂ Cl _{2 was added} to improve the solubility and the mixture was again heated at 45 °C. The mixture was cooled to room temperature and water was added dropwise. The resulting mixture was stirred at room temperature overnight and then extracted with CH _{2 2} Cl. The organic extracts were concentrated and the residue was triturated with EtOAc. The solid was filtered, washed with diethyl ether and dried in a vacuum oven for several hours to afford 6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5- (1) '-Methyl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-oxirane-1, 6-Dihydro-pyridazin-3-yl]-phenyl}-2H-phthalazin-1-one (100 mg, 55%). NMR confirmed the presence of a small amount of isopropyl acetate. LC/MS observed [M+H] ⁺ 606.1.

Example 3

2-{3-[5-(5-azetidin-1-ylmethyl-1-methyl-1H-pyrazol-3-ylamino)-1-methyl-6-sideoxy -1,6-dihydro-pyridazin-3-yl]-2-hydroxymethyl-phenyl}-6-tert-butyl-2H-phthalazin-1-one

Purging 2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[-acetic acid] in a microwave vial with nitrogen 1,3,2]diboron 2-yl)-benzyl ester (which can be prepared as described in Example 5; about 75% purity; 206 mg, about 0.32 mmol), 4-(5-(azetidin-1-ylmethyl) 1-methyl-1H-pyrazol-3-ylamino)-6-chloro-2-methylpyridazin-3(2H)-one (which can be as described in Berthel, SJ et al, US 20120040949, Example 58 Prepared as described in the intermediate; 89 mg, 0.29 mmol), bis(dibenzylideneacetone)palladium (Aldrich; 8.3 mg, 14 μmol), XPhos (2-(dicyclohexylphosphino)-2', 4 a mixture of ',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 13.7 mg, 29 μmol) and tripotassium phosphate (184 mg, 0.87 mmol) with degassed n-butanol (2.3 the mixture mL) and water (576 μL) the mixture was heated at 115 deg.] C overnight in a sand bath. the mixture was evaporated and purified by chromatography to give (with 0-25% MeOH / CH ₂ Cl ₂ fractions) as a yellow oil (150 mg), MeOH (2.4 mL) and EtOAc (EtOAc (EtOAc)EtOAc. Dry overnight in a vacuum oven to give 2-{3-[5-(5-azetidin-1-ylmethyl-1-methyl) as an off-white solid. -1H-pyrazol-3-ylamino)-1-methyl-6-yloxy-1,6-dihydro-pyridazin-3-yl]-2-hydroxymethyl-phenyl}-6 - tert-butyl -2H- phthalazin-1-one (72 mg, 43%) LC / MS observations [M ⁺ H] + 581.4.

Example 4

6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((S)-1-methyl-pyrrolidin-2-yl)-pyridine-2 -ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one

Purging 2-(6-t-butyl-1-oxooxy-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1, 3,2]diboron 2-yl)-benzyl ester (which can be prepared as described above; about 75% purity; 172 mg, about 0.27 mmol), (S)-6-chloro-2-methyl-4-(5-( 1-Methylpyrrolidin-2-yl)pyridin-2-ylamino)pyridazine-3(2H)-one (which can be prepared as described in Berthel, SJ et al, US 20120040949, Example 18, Step 3; 77 Mg, 0.24 mmol), bis(dibenzylideneacetone)palladium (Aldrich; 6.9 mg, 12 μmol), XPhos (2-(dicyclohexylphosphino)-2', 4',6'-triisopropyl a mixture of keto-1,1'-biphenyl, Strem Chemicals; 14.3 mg, 30 μmol) and tripotassium phosphate (153 mg, 0.72 mmol) with degassed n-butanol (2.4 mL) and water (600 μL) a mixture. The mixture was heated in a sand bath at 115 ° C for 2.5 hours. Diatomaceous earth mixture was filtered, concentrated and purified by chromatography (with 0-25% MeOH / CH ₂ Cl ₂ fractions), to give 100 mg of the product mixture. MeOH (1.58 mL) and CH ₂ Cl _{2 were} added sequentially to dissolve the solid. Potassium carbonate (65.4 mg, 0.47 mmol) was added and the mixture was heated at 44 ° C for 1.5 h. Water was added dropwise and the mixture was stirred at room temperature for 2 hours. The mixture was filtered and the solid was washed with water and Et ₂ O and dried for a weekend in vacuo to afford an off-white crystalline solid of 6- tert-butyl-2- (2-hydroxymethyl-3- {1-methyl-5- [ 5-((S)-1-methyl-pyrrolidin-2-yl)-pyridin-2-ylamino]-6-oxooxy-1,6-dihydro-pyridazin-3-yl}- Phenyl)-2H-phthalazin-1-one (85 mg, 91%). [M+H] ⁺ 592.5.

2-(6-T-butyl-1-oxo-1H-pyridazin-2-yl)-6-{1-methyl-5-[5-((S)-1-methyl-) Pyrrrolidin-3-yl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenylmethyl ester

6-Chloro-2-methyl-4-[5-((S)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-2H-pyridazin-3-one ( It can be prepared as described in Berthel, SJ et al, US20120040949, Example 32, Step 4; 150 mg, 469 μmol), bispinacol borate (155 mg, 610 μmol), potassium acetate (138 mg, 1.41) Methyl) and dioxane (7 mL) were degassed under argon. Add XPhos (2-(dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 33.5 mg, 70.4 μmol) and palladium(II) acetate (5.3 mg, 23.5 μmol) and the reaction mixture was stirred under argon at 100 ° C (external temperature) for 30 minutes. The flask was taken out of the heating bath and 2-(6-t-butyl-1-oxo-oxazine-2(1H)-yl)-6-chlorobenzyl acetate (Example 4; 199 mg, 516 μmol) was added. ), potassium carbonate (194 mg, 1.41 mmol), tricyclohexylphosphine (13.2 mg, 46.9 μmol), and bis(dibenzylideneacetone)palladium(0) (13.5 mg, 23.5 μmol) and water (1.5 ml) . The reaction mixture was flushed with argon and heated at 80 ° C for 7 hours with vigorous stirring. The reaction mixture was cooled to room temperature and poured into water and EtOAc. The EtOAc layer was separated and aqueous was extracted withEtOAc. Combined organic layers were washed with brine, dried (Na ₂ SO _4), filtered and evaporated. The residue was purified by a 12 g column (solvent eluting with CH ₂ Cl ₂ /MeOH/NH ₄ OH 89:9:2->66:27:7) using the Analogix system to give 2-(6-t-butyl acetate) -1-Sideoxy-1H-pyridazin-2-yl)-6-{1-methyl-5-[5-((S)-1-methyl-pyrrolidin-3-yl)-pyridine- 2-Aminoamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenylmethyl ester (78 mg, 26%). LC/MS observed [M+H] ⁺ 634.

Example 5

6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((S)-1-methyl-pyrrolidin-3-yl)-pyridine-2 -ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one

2-(6-Tert-butyl-1-oxo-1H-pyridazin-2-yl)-6-{1-methyl-5-[5-((S)-1-methyl)acetate -pyrrolidin-3-yl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenylmethyl ester (78 mg, 0.124 mmol), A mixture of K ₂ CO ₃ (34.2 mg, 0.247 mmol) and MeOH (10 mL) was stirred at 40 ° C for 75 min and then cooled to room temperature. Water and CH ₂ Cl _{2 were added} and the aqueous layer was extracted with CH ₂ Cl ₂ . Dried ₍₂ SO ₄ Na) combined organic layers were, filtered and evaporated. The residue was triturated with EtOAc ₍ EtOAc) EtOAc (EtOAc) ((S)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl -2H-phthalazin-1-one (57 mg, 79%). LC/MS observed [M+H] ⁺ 592.

2-(6-Tertibutyl-1-oxo-1H-pyridazin-2-yl)-6-{1-methyl-5-[5-((R)-1-methyl-pyrrole Pyridin-3-yl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-benzaldehyde

6-Chloro-2-methyl-4-[5-((R)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-2H-pyridazin-3-one ( It can be prepared as described in Berthel, SJ et al., US20120040949, Example 33, Step 1; 150 mg, 469 μmol), bis-pinacol borate (155 mg, 610 μmol), potassium acetate (138 mg, 1.41). Methyl) and dioxane (7 mL) were degassed under argon. Add XPhos (2-(dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 33.5 mg, 70.4 μmol) and palladium(II) acetate (5.3 mg, 23.5 μmol) and the reaction mixture was stirred under argon at 100 ° C (external temperature) for 30 minutes. The flask was taken out of the heating bath and 2-(6-t-butyl-1-oxo-oxazine-2(1H)-yl)-6-chlorobenzaldehyde (Example 2; 160 mg, 469 μmol) was added. Potassium carbonate (194 mg, 1.41 mmol), tricyclohexylphosphine (13.2 mg, 46.9 μmol), and bis(dibenzylideneacetone)palladium(0) (13.5 mg, 23.5 μmol) and water (1.5 ml). The reaction mixture was flushed with argon and heated at 80 ° C for 1 hour with vigorous stirring. The reaction mixture was cooled to room temperature and poured into water and EtOAc. The EtOAc layer was separated and aqueous was extracted withEtOAc. Combined organic layers were washed with brine, dried (Na ₂ SO _4), filtered and evaporated. The residue was purified on a Supelco 24 G column (solvent eluting with CH ₂ Cl ₂ /MeOH/NH ₄ OH 77:18:5) using the Analogix system to give 2-(6-t-butyl-1- oxoacetate -1H-pyridazin-2-yl)-6-{1-methyl-5-[5-((R)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino] -6-Phenoxy-1,6-dihydro-pyridazin-3-yl}-phenylmethyl ester (16 mg, 6%). LC/MS observed [M+H] ⁺ 633.9.

Example 6

6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((R)-1-methyl-pyrrolidin-3-yl)-pyridine-2 -ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one Preparation of 6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((R)-1-methyl-pyrrolidin-3-yl)-pyridine- 2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one

2-(6-Tert-butyl-1-oxo-1H-pyridazin-2-yl)-6-{1-methyl-5-[5-((R)-1-methyl-)-acetic acid Pyrrrolidin-3-yl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenylmethyl ester (16 mg, 27 μmol), boron A mixture of sodium hydride (1.64 mg, 43.4 μmol) and methanol (1 mL) was stirred under nitrogen for 30 min. Add aqueous saturated NH ₄ Cl and the mixture was stirred for 5 minutes and then extracted twice with CH _{2 2} Cl. The combined organic layers were dried, filtered and evaporated. By column chromatography _{(CH 2 Cl 2 / MeOH / NH} 4 OH 89: 9: 2-> 66: 27: 7 eluting) and the residue was purified product was vacuum oven dried at 50 deg.] C overnight to afford 6-Terbutyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((R)-1-methyl-pyrrolidin-3-yl) as a pale yellow solid )-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one (11.8 mg, 74%) ). LC/MS observed [M+H] ⁺ 592.

2-(2-{2-Mercapto-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro- [3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-sidedoxy-1,2 -dihydro-pyridazin-6-yl)-2-methyl-propanenitrile

Solution A was prepared as follows: 6-chloro-2-methyl-4-(5-(1-methylpiperidin-4-yl)pyridin-2-ylamino)pyridazine-3(2H)-one (This can be prepared as described in Berthel, SJ et al., US20120040949, Step 2 of Process I-67; 125 mg, 0.374 mmol), XPhos (2-(dicyclohexylphosphino)-2', 4', 6 '-Triisopropyl-1,1'-biphenyl, Strem Chemicals; 27 mg, 56 μmol), potassium acetate (110 mg, 1.1 mmol), bis-pinacol borate (124 mg, 0.49 mmol) A mixture of dioxane (4.4 mL) was vacuum degassed under reduced pressure and placed under argon. Palladium(II) acetate (9.25 mg, 41.2 μmol) was added at room temperature and the vacuum degassing cycle was repeated. The reaction mixture was stirred at 100 ° C (external temperature) under argon for 16 minutes. The reaction mixture was cooled to ambient temperature and filtered through celite pad with argon gas (washing celite with additional 2.5 mL of dioxane) to give solution A.

Solution B was prepared as follows: 2-(2-(3-bromo-2-methylindenylphenyl)-1-yloxy-1,2-dihydropyridazin-6-yl)-2-methyl Propionitrile (Example 8; 158 mg, 399 μmol), tricyclohexylphosphine (34.2 mg, 122 μmol), K ₂ CO ₃ (276 mg, 2 mmol), n-BuOH (486 μL), water (1.52 mL) The mixture with dioxane (2.03 mL) was degassed. Bis(dibenzylideneacetone)palladium(0) (34.4 mg, 59.8 μmol) was added at room temperature. The mixture was again degassed under vacuum and then heated in a heating bath set at 110 °C.

Solution A was added to Solution B and the resulting mixture was stirred at 110 ° C for 35 minutes. Heating was stopped and the reaction mixture was stirred overnight. The reaction mixture was filtered over EtOAc (EtOAc)EtOAc. EtOAc (30 mL) and water (30 mL) were added to the filtrate and the filtrate was shaken in a separating funnel. The EtOAc phase was collected. The aqueous layer was combined organic layers were back-extracted with EtOAc (2 × 20 mL) and dried (Na ₂ SO _4), filtered and evaporated. SF15-24g system by using an Analogix column (/ CH ₂ Cl ₂ eluting with 3% -18% MeOH) and the residue was purified by chromatography, to give a gray powder of 2- (2- {2-acyl -3 -[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamine -6-Sideoxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-yloxy-1,2-dihydro-pyridazin-6-yl)-2 -Methyl-propionitrile (178 mg, 73%). LC/MS observed [M+H] ⁺ 615.

Example 7 2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro- [3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-sidedoxy-1,2 -dihydro-pyridazin-6-yl)-2-methyl-propanenitrile

2-(2-{2-Mercapto-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro) -[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-yloxy-1, 2- dihydro - phthalazin-6-yl) -2-methyl - propionitrile (example 17; 178 mg, 0.29 mmol) was dissolved in a mixture of MeOH (4 mL) and CH ₂ Cl ₂ (2.3 mL) and the Cool to 0 °C. Was added dropwise _{NaBH 4 (55 mg, 1.5 mmol} ) in water (0.5 mL) of the solution and the mixture was stirred for 10 minutes. Additional NaBH ₄ (55 mg, 1.5 mmol) in water (0.5 mL) was then added and the mixture was stirred for further 10 min. Water (60 mL) and CH ₂ Cl ₂ (60 mL) were added and the material was shaken in a sep. funnel. The organic layer was collected and washed with 50% saline solution and the aqueous phase diluted _{CH 2 Cl 2 (2 × 40} mL) and extracted with trans. Combined organic layers were dried (MgSO _4), filtered and evaporated. By using an Analogix purification system SF15-12G silicon dioxide column (3% -14% MeOH / CH 2 Cl 2) Purification of the residue, to give a pale yellow solid (138 mg). This material was recrystallized from CH ₂ Cl ₂ /hexane to give 2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1') as white crystals. , 2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazine -3-yl]-phenyl}-1-o-oxy-1,2-dihydro-pyridazin-6-yl)-2-methyl-propanenitrile (109 mg, 61%). LC/MS observed [M+H] ⁺ 617.

Example 8

6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(5-methyl-4,5,6,7-tetrahydro-pyrazolo[1,5 -a]pyrazin-2-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-2H-pyridazin-1-one

Adding a mixture of degassed n-BuOH (4 mL) and water (1 mL) to a vial of 2-(6-t-butyl-1-one-oxyl-1H-pyridazin-2-yl) )-6-(4,4,5,5-tetramethyl-[1,3,2]diboron 2-yl)-benzyl ester (which can be prepared as described in Example 5; about 75% purity; 364 mg, about 0.57 mmol), 6-chloro-2-methyl-4-(5-methyl- 4,5,6,7-tetrahydro-pyrazolo[1,5-a]pyrazin-2-ylamino)-2H-pyridazin-3-one (which can be as Berthel, SJ et al, US20120040949) Prepared as described in the preparation of Example 59; 150 mg, 0.51 mmol), bis(dibenzylideneacetone)palladium (Aldrich; 14.6 mg, 25 μmol), XPhos (2-(dicyclohexylphosphino) a mixture of -2', 4', 6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 24.3 mg, 51 μmol) and tripotassium phosphate (324 mg, 1.5 mmol). The vial was capped and the mixture was heated in a sand bath at 115 °C for 2.5 hours. Diatomaceous earth mixture was filtered, evaporated and purified by chromatography (0-25% MeOH / CH ₂ Cl ₂ fractions), to give 175 mg of the product mixture. MeOH (2.9 mL) and potassium carbonate (119 mg, 0.86 mmol) were added and the mixture was stirred at 40 ° C for one hour. Water was added dropwise and the resulting mixture was stirred at room temperature for 1 hour. Water was added dropwise and the mixture was stirred at room temperature for 1 hour. The resulting solid was filtered and dried in vacuo to give 6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(5-methyl) as a white crystalline solid. -4,5,6,7-tetrahydro-pyrazolo[1,5-a]pyrazin-2-ylamino)-6-o-oxy-1,6-dihydro-pyridazine-3- Base]-phenyl}-2H-phthalazin-1-one (147 mg, 51%). LC/MS observed [M+H] ⁺ 567.0.

Example 9

6-Tertibutyl-2-{3-[5-(1'-ethyl-1',2',3',4',5',6'-hexahydro-[3,4'] Pyridine-6-ylamino)-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl]-2-hydroxymethyl-phenyl}-2H-pyridazine- 1-ketone

Argon was bubbled through 6-chloro-4-(1'-ethyl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridine-6 -ylamino)-2-methyl-2H-pyridazin-3-one (which can be prepared as described in Berthel, SJ et al, US 20120040949, Example 69 intermediate; 133 mg, 0.38 mmol), acetic acid 2 -(6-t-butyl-1-oxo-oxazine-2(1H)-yl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) benzyl ester (which can be prepared as described in Example 5; about 75% purity; 242 mg, 381 μmol) and tripotassium phosphate (202 mg, 953 μmol), water (1.25 mL), and nBuOH ( Mix 5 mL) for 5 minutes. Add XPhos (2-(dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 18.2 mg, 38 μmol) and bis(diphenylene) Methylacetone) palladium (0) (Strem Chemicals; 11.0 mg, 19.1 μmol). The tube was sealed and the reaction mixture was heated at 115 °C for 6 hours. The reaction mixture was poured into water (100 mL)EtOAc. Combined organic layers were washed with brine, dried (MgSO _4), filtered and evaporated. The residue was purified by chromatography (sequentially with 0-6% MeOH / CH ₂ Cl ₂ and _{NH 4 OH / MeOH / CH 2} Cl 2 4:14:82 fractions) and then purified with EtOAc / Et ₂ O triturated to give 6-tert-butyl-2-{3-[5-(1'-ethyl-1',2',3',4',5',6'-hexahydro-[3] , 4']bipyridin-6-ylamino)-1-methyl-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-2-hydroxymethyl-phenyl}- 2H-phthalazin-1-one (32 mg, 13%). ¹ H NMR confirmed the presence of <3% impurity, 2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6-[5-(1'-ethyl-1) acetate ',2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-1-methyl-6-oxirane-1,6 -Dihydro-pyridazin-3-yl]-phenylmethyl ester. LC/MS observed [M+H] ⁺ 620.1.

Example 10

6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(morpholin-4-carbonyl)-pyridin-2-ylamino]-6-side Oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one

Adding a degassed mixture containing n-BuOH (2.29 mL) and water (0.57 mL) to the acetic acid 2-(6-t-butyl-1-one-oxy-1H-pyridazin-2-yl) in a microwave vial )-6-(4,4,5,5-tetramethyl-[1,3,2]diboron 2-yl)-benzyl ester (which can be prepared as described in Example 5; about 75% purity; 204 mg, about 0.32 mmol), 6-chloro-2-methyl-4-[5-(morpholine) 4-carbonyl)-pyridin-2-ylamino]-2H-pyridazin-3-one (which can be prepared as described in Berthel, SJ et al., US 20120040949, Process I-2; 100 mg, 0.29 mmol) , bis(diphenylpyrimidinone)palladium (8.2 mg, 14.3 μmol), XPhos (2-(dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-linked A mixture of benzene, Strem Chemicals; 13.6 mg, 28.6 μmol) and tripotassium phosphate (152 mg, 0.72 mmol). The mixture was heated overnight at 115 ° C in a sand bath. Diatomaceous earth mixture was filtered, evaporated and purified by chromatography (0-20% MeOH / CH ₂ Cl ₂ fractions), to give a solid. Methanol was added and the mixture was filtered and dried in a vacuum oven for one weekend to give 6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-( Morpholine-4-carbonyl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-pyridazin-1-one (46 mg, 26%). LC/MS observed [M+H] ⁺ 622.4.

2-[2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1 -methyl-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-2-carboxanyl-phenyl)-1-yloxy-1,2-dihydro-pyridazine -6-yl]-2-methyl-propionitrile

Solution A was prepared as follows: 4-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-6-chloro -2-Methyl-2H-pyridazin-3-one (which can be prepared as described in Berthel, SJ et al., US20120040949, I-13, Method 2; 150 mg, 0.41 mmol), XPhos (2-( Dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 29.5 mg, 62 μmol), potassium acetate (121 mg, 1.2 mmol), double A mixture of terbibenzoic acid borate (Matrix Scientific; 136 mg, 0.54 mmol) and dioxane (5.3 mL) was vacuum degassed under reduced pressure and placed under argon atmosphere. Palladium(II) acetate (10.2 mg, 45 μmol) was added at room temperature and the vacuum degassing cycle was repeated. The reaction mixture was stirred at 100 ° C (oil bath temperature) under argon for 16 minutes. The reaction mixture was filtered through a pad of diatomaceous earth using argon gas (washing diatomaceous earth with 2.5 mL of dioxane) to give solution A.

Solution B was prepared as follows: 2-(2-(3-bromo-2-methylindenylphenyl)-1-yloxy-1,2-dihydropyridazin-6-yl)-2-methyl Propionitrile (Example 8; 156 mg, 394 μmol), Tricyclohexylphosphine (33.8 mg, 120 μmol), K ₂ CO ₃ (276 mg, 2 mmol), n-BuOH (480 μL), water (1.5 mL) A mixture of dioxane (2 mL) was degassed. Bis(diphenylmethyleneacetone)palladium(0) (34 mg, 59 μmol) was added at room temperature. The mixture was again degassed under vacuum and then heated in an oil bath set at 110 °C.

Solution A was added to Solution B and the resulting mixture was stirred at 110 ° C for 60 minutes. The reaction mixture was cooled to room temperature and filtered over EtOAc. The diatomaceous earth was washed with EtOAc. EtOAc (30 mL) and water (30 mL) were added toEtOAc. The aqueous layer was combined organic layers were back-extracted with EtOAc (2 × 20 mL) and dried (Na ₂ SO _4), filtered and evaporated. SF15-12g system by using an Analogix column chromatography (/ CH ₂ Cl ₂ eluting with 3% -18% MeOH) The residue was purified to give a pale brown powder of 2- [2- (3- {5- [ 5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl-6-oxirane-1, 6-Dihydro-pyridazin-3-yl}-2-carboxanyl-phenyl)-1-yloxy-1,2-dihydro-pyridazin-6-yl]-2-methyl-propane Nitrile (201 mg, 79%). LC/MS observed [M+H] ⁺ 645.0.

Example 11

2-[2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1 -methyl-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-1-yloxy-1,2-dihydro-pyridazine -6-yl]-2-methyl-propionitrile

To 2-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl-6 -Sideoxy-1,6-dihydro-pyridazin-3-yl}-6-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-benzaldehyde (201 mg, 0.31 mmol) in MeOH (4.5 mL) and in the CH ₂ Cl ₂ (2.3 mL) the mixture was dropwise added _{NaBH 4 (59 mg, 1.6 mmol} ) in water (0.5 mL) in the solution and at 0 ℃ The resulting mixture was stirred for 10 minutes. Add _{NaBH 4 (59 mg, 1.6 mmol} ) in water (0.5 mL) of the solution and the mixture was further stirred for 10 minutes. Water (60 mL) and CH ₂ Cl ₂ (60 mL) were added and the layers were separated. The organic layer was extracted with 50% anti-saline solution (60 mL) were combined and washed with an aqueous solution of _{CH 2 Cl 2 (2 × 40} mL). Combined organic layers were dried (MgSO _4), filtered and evaporated. SF15-12G by using an Analogix purification system column (with a 2-14% MeOH / CH ₂ Cl ₂ fractions) to give the residue, followed by self-CH ₂ Cl ₂ / hexane and then crystallized to give white crystals of 2- [ 2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl -6-Phenoxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-1-yloxy-1,2-dihydro-pyridazine-6- 2-methyl-propionitrile (145 mg, 72%). LC/MS observed [M+H] ⁺ 647.1.

2-(6-T-butyl-1-oxo-1H-pyridazin-2-yl)-6-(5-{5-[2-(3-hydroxy-propylamino)-1) acetate ,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl)-phenylmethyl ester

Rinse 4-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-6-chloro-2- with argon Methyl-2H-pyridazin-3-one (which can be prepared as described in Berthel, SJ et al., US20120040949, Method 2, Process I-13; 1.50 g, 4.12 mmol), 2-(6-third) Butyl-1-oxooxy-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaboron -2-yl)-benzyl ester (ELN003987-149; 83% purity; 2.95 g, 5.14 mmol), bis(dibenzylideneacetone)palladium (Strem Chemicals; 119 mg, 0.21 mmol), XPhos (2- (dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 197 mg, 0.41 mmol) and tripotassium phosphate (2.19 g, 10.3 mmol) A mixture of nBuOH (33 mL) and water (8.25 mL) was then heated overnight at 100 °C. The mixture was filtered through celite and concentrated. The residue was purified (:: 144) eluting _{2 Cl 2 / MeOH / NH 4} OH (82 with CH ₂ Cl ₂ to CH) using Analogix purification system. The pump of the purification system began to fail during the purification process. Using different Analogix purification system (using _{CH 2 Cl 2 / MeOH / NH} 4 OH (82: 14: 4-> 55: 36: 9) as eluent) eluting the product from the column. The product was slurried/dissolved in Et ₂ O / EtOAc (2: 1) and the mixture was stood for 48 hr. The solid was filtered to give 2-(6-t-butyl-1-ylidene-1H-pyridazin-2-yl)-6-(5-{5-[2-(3). -hydroxy-propylamino)-1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxirane-1,6-dihydro-indole Pyrazin-3-yl)-phenylmethyl ester (525 mg, 19%). LC/MS observed [M+H] ⁺ 696.0.

Example 12

6-Terbutyl-2-[2-hydroxymethyl-3-(5-{5-[2-(3-hydroxy-propylamino)-1,1-dimethyl-ethoxy] -pyridin-2-ylamino}-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl)-phenyl]-2H-phthalazin-1-one

2-(6-T-butyl-1-oxo-1H-pyridazin-2-yl)-6-(5-{5-[2-(3-hydroxy-propylamino)-1) acetate ,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl)-phenylmethyl ester (example 24; 295 mg, 0.42 mmol) , the mixture (117 mg, 0.85 mmol) and MeOH (30 mL) of K ₂ CO ₃ was stirred at 40 ℃ 75 minutes and then cooled to room temperature. Water and CH ₂ Cl _{2 were added} and the aqueous layer was extracted with CH ₂ Cl ₂ . Dried ₍₂ SO ₄ Na) combined organic layers were, filtered and evaporated. Using 12 g Silicycle column (with _{1% -15% MeOH / CH 2} Cl 2 fractions) by the residue purified by chromatography. A homogeneous sample of the product was evaporated and the residue was dried in vacuo at 50 ° C to afford 6-t-butyl-2-[2-hydroxymethyl-3-(5-{5-[2 -(3-hydroxy-propylamino)-1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxirane-1,6-di Hydrogen-pyridazin-3-yl)-phenyl]-2H-phthalazin-1-one (119 mg, 43%). LC/MS observed [M+H] ⁺ 654.2.

Example 13

6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(1'-oxetan-3-yl-1',2',3',4 ',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl }-2H-pyridazin-1-one

Argon was bubbled through 6-chloro-2-methyl-4-(1'-oxetan-3-yl-1',2',3',4',5',6'-hexa Hydrogen-[3,4']bipyridin-6-ylamino)-2H-pyridazin-3-one (which can be prepared as described in Berthel, SJ et al, US 20120040949, Example 80, Step 1; 213 mg, 567 μmol), 2-(6-t-butyl-1-oxo-1H-pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1,3] , 2] boron trioxide 2-yl)-benzyl ester (which can be prepared as described in Example 5; about 75% purity; 648 mg, about 1.0 mmol) and tripotassium phosphate (301 mg, 1.42 mmol), water (1.25 mL) and A mixture of nBuOH (5 mL) was used for 5 minutes. Add XPhos (2-(dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, Strem Chemicals; 27 mg, 57 μmol) and bis(diphenylene) Methylacetone) palladium (0) (Strem Chemicals; 16.3 mg, 28 μmol). The tube was sealed and the reaction mixture was heated at 115 °C for 4 hours. The reaction mixture was poured into water (100 mL) and the mixture (5 × 75 mL) and extracted with CH ₂ Cl _2. Combined organic layers were washed with brine, dried (Na ₂ SO _4), filtered and evaporated. By chromatography (0-4% MeOH / CH ₂ Cl ₂ fractions) to give the residue, to give a mixture of ethanol and acetate the product (374 mg). THF (12 mL) and 1 N NaOH (2 mL) were added and the mixture was warmed overnight at 60 °C. The mixture was poured into water and extracted three times with EtOAc and _extracted once with CH ₂ Cl. The organic extracts were washed with brine, dried (Na ₂ SO _4), filtered and evaporated. By chromatography (eluted with _{1% -5% MeOH / CH 2} Cl 2 fractions) to give the residue, to give a glass. Add EtOAc (4 mL) and Et ₂ O (30 mL) and the white precipitate was filtered off and washed with ether to give a white powder of 6- tert-butyl-2- {2-hydroxy-3- [ 1-Methyl-5-(1'-oxetan-3-yl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridine -6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-2H-phthalazin-1-one (246 mg, 67%). LC/MS observed [M+H] ⁺ 648.1.

6-Chloro-4-{5-[2-(3,3-difluoro-azetidin-1-yl)-1,1-dimethyl-ethoxy]-pyridin-2-ylamine -2--methyl-2H-pyridazin-3-one

2-[6-(6-Chloro-2-methyl-3-oxo-2,3-dihydro-pyridazin-4-ylamino)-pyridin-3-yloxy]-2-yl Base-propionaldehyde (which can be prepared as described in Berthel, SJ et al., US 20120040949, Process 1 of Process I-13; 665 mg, 2.06 mmol), 3,3-difluoroazetidine hydrochloride ( A mixture of 347 mg, 2.68 mmol) and 1,2-dichloroethane (70 mL) was stirred at room temperature for 1 hour. Sodium triethoxysulfonate (655 mg, 3.09 mmol) was added and the mixture was stirred at room temperature for 18 h. Saturated aqueous NaHCO ₃ was added and the mixture (5 × 20 mL) and extracted with CH ₂ Cl _2. Combined organic layers were dried (MgSO _4), filtered and evaporated. The residue was purified by chromatography (30% - 50%EtOAcEtOAcEtOAc) 1-yl)-1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-2-methyl-2H-pyridazin-3-one (566 mg, 69%). LC/MS observed [M+H] ⁺ 400.0.

Example 14

6-t-butyl-2-[3-(5-{5-[2-(3,3-difluoro-azetidin-1-yl)-1,1-dimethyl-ethoxy ]]-pyridin-2-ylamino}}-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl)-2-hydroxymethyl-phenyl]-2H- Pyridazine-1-one

6-Chloro-4-{5-[2-(3,3-difluoro-azetidin-1-yl)-1,1-dimethyl-ethoxy]-pyridine by vacuuming -2-ylamino}-2-methyl-2H-pyridazin-3-one (Example 27; 120 mg, 0.3 mmol), 2-(6-tert-butyl-1-oxo-1H acetate -pyridazin-2-yl)-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaboron 2-yl)-benzyl ester (which can be prepared as described in Example 5; 310 mg, 0.39 mmol), tripotassium phosphate (140 mg, 0.66 mmol), nBuOH (4 mL), and water (1 mL) The mixture was degassed and backfilled with argon. Add XPhos (2-(dicyclohexylphosphino)-2',4',6'-triisopropyl-1,1'-biphenyl, available from Strem Chemicals; 21.5 mg, 45 μmol) and double ( Diphenylmethyleneacetone) palladium (0) (available from Strem Chemicals; 12.1 mg, 21 μmol). The mixture was again degassed and then heated at 115 ° C for 3 hours. Water (35 mL) and EtOAc (35 mL) were added. The EtOAc layer was washed with brine (35 mL Dried ₍₂ SO ₄ Na) combined organic layers were, filtered and evaporated. The residue was purified by EtOAc (EtOAc:EtOAc) -difluoro-azetidin-1-yl)-1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxirane-1, 6-Dihydro-pyridazin-3-yl)-2-hydroxymethyl-phenyl]-2H-phthalazin-1-one (130 mg, 65%). LC/MS observed [M+H] ⁺ 672.0.

2-(2-{2-Mercapto-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro- [3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-sidedoxy-1,2 ,3,4-tetrahydro-isoquinolin-6-yl)-2-methyl-propanenitrile

6-Chloro-2-methyl-4-(5-(1-methylpiperidin-4-yl)pyridin-2-ylamino)pyridazine-3(2H)-one by vacuuming It can be prepared as described in Berthel, SJ et al., US20120040949, Step 2 of Process I-67; 140 mg, 0.42 mmol), XPhos (2-(dicyclohexylphosphino)-2', 4', 6'- Triisopropyl-1,1'-biphenyl, Strem Chemicals; 30 mg, 63 μmol), potassium acetate (123 mg, 1.26 mmol), bis-pinacol borate (Matrix Scientific; 138 mg, 0.55 mmol The mixture with dioxane (7 mL) was degassed and then backfilled with argon. Palladium(II) acetate (10.4 mg, 46 μmol) was added and the mixture was again degassed. The mixture was heated at 100 ° C for 16 minutes under argon. Using argon pressure through the diatomaceous earth filler The reaction mixture was filtered (washing diatomaceous earth with an additional 2.5 mL of dioxane) to give solution A.

Solution B was prepared as follows: 2-[2-(3-bromo-2-methylindolyl-phenyl)-1-yloxy-1,2,3,4-tetrahydro-isoquinoline-6- 2-methyl-propionitrile (which can be prepared as described in Berthel, SJ et al, US20100222325, Example 153; 166 mg, 0.42 mmol), tricyclohexylphosphine (36 mg, 128 μmol), K ₂ A mixture of CO ₃ (290 mg, 2.1 mmol), n-BuOH (534 μL), water (2.1 mL) and dioxane (2.22 mL) was degassed. Bis(diphenylmethyleneacetone)palladium(0) (36.1 mg, 63 μmol) was added at room temperature. The mixture was again degassed under vacuum and then heated in a heating bath set at 110 °C.

Solution A was added to Solution B and the resulting mixture was stirred at 110 ° C for 60 minutes. Heating was stopped and the reaction mixture was stirred overnight. The reaction mixture was filtered over EtOAc (EtOAc)EtOAc. EtOAc (30 mL) and water (30 mL) were added toEtOAc. The aqueous layer was combined organic layers were back-extracted with EtOAc (2 × 20 mL) and dried (Na ₂ SO _4), filtered and evaporated. SF25-24g system by using an Analogix column chromatography (using _{2% -13% MeOH / CH 2} Cl 2 fractions) to give the residue, to give a light brown semi-solid of 2- (2- {2-acyl -3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridine-6- Amino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-sidedoxy-1,2,3,4-tetrahydro-isoquinoline -6-yl)-2-methyl-propionitrile (158 mg, 61%). LC/MS observed [M+H] ⁺ 616.0.

Example 15

2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6'-hexahydro- [3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-sidedoxy-1,2 ,3,4-tetrahydro-isoquinolin-6-yl)-2-methyl-propanenitrile

A solution of NaBH ₄ (49 mg, 1.3 mmol) in water (0.75 mL) was added to cooled (~0 °C) 2-(2-{2-carbazyl-3-[1-methyl-5 -(1'-methyl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-sideoxy -1,6-dihydro-pyridazin-3-yl]-phenyl}-1-o-oxy-1,2,3,4-tetrahydro-isoquinolin-6-yl)-2-methyl - a mixture of propionitrile (158 mg, 0.26 mmol), CH 2 Cl 2 (2 mL) and MeOH (2 mL) of the. The mixture was stirred for 10 minutes. Add _{NaBH 4 (49 mg, 1.3 mmol} ) in water (0.75 mL) in the solution and the mixture was stirred for 10 minutes. Add _{NaBH 4 (66 mg, 1.7 mmol} ) in water (0.75 mL) in the solution and the mixture was stirred for 5 minutes. The upper aqueous layer was removed and MeOH (1 mL) was added. Two additional portions of NaBH ₄ (66 mg, 1.7 mL) in water (0.75 mL) were added at 10 min intervals and the reaction mixture was stirred for 10 min. Water (60 mL) and _{CH 2 Cl 2 (50 mL)} . The mixture was shaken in a separatory funnel and the organic layer was separated. The organic layer was washed with 50% brine solution _{(50 mL) CH 2 Cl 2} (2 × 40 mL) were combined and back-extracted water phase was washed with. Combined organic layers were dried (MgSO _4), filtered and evaporated. SF15-12G by using an Analogix purification system column (with _{1% -16% MeOH / CH 2} Cl 2 fractions) to give the residue, to give an off-white solid (101 mg). This material is recrystallized from CH ₂ Cl ₂ /hexane to give 2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1') as an off-white powder. , 2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazine 3-yl]-phenyl}-1-o-oxy-1,2,3,4-tetrahydro-isoquinolin-6-yl)-2-methyl-propanenitrile (81 mg, 92%) . LC/MS observed [M+H] ⁺ 618.2.

Biological example Bruton's tyrosine kinase (Btk) inhibition assay

This assay captures the radioactive ³³ P phosphorylated product via filtration. The interaction of Btk, biotinylated SH ₂ peptide receptor (Src homolog) with ATP causes the peptide to undergo phosphorylation. The biotinylated product is the bound streptavidin agarose beads. The combined radiolabeled product is detected by a scintillation counter.

The assay plates were 96-well polypropylene (Greiner) and 96-well 1.2 μm hydrophilic PVDF filter disc (Millipore). The concentrations reported here are the final assay concentrations: 10-100 μM compound in DMSO (Burdick and Jackson), 5-10 nM Btk enzyme (His-labeled, full-length), 30 μM peptide receptor (Biotin-Aca-AAAEEIYGEI- NH ₂ ), 100 μM ATP (Sigma), 8 mM imidazole (Sigma, pH 7.2), 8 mM glycerol-2-phosphate (Sigma), 200 μM EGTA (Roche Diagnostics), 1 mM MnCl ₂ (Sigma), 20 mM MgCl ₂ (Sigma), 0.1 mg/ml BSA (Sigma), 2 mM DTT (Sigma), 1 μCi ³³ P ATP (Amersham), 20% streptavidin agarose beads (Amersham), 50 mM EDTA (Gibco), 2 M NaCl (Gibco), 2 M NaCl + 1% phosphoric acid (Gibco), Microscint-20 (Perkin Elmer).

Use the information generated by the analysis of a standard 96 well plate template, since 10 data points measured value ₅₀ calculated for each compound IC. Each plate was tested for one control compound and seven unknown inhibitors, and each plate was performed twice. Typically, the compound is diluted in a half log, starting at 100 μM and terminating at 3 nM. The control compound was staurosporine. The background was counted in the absence of peptide acceptor. Total activity was determined in the presence of peptide acceptors. Btk inhibition was determined using the following protocol.

1) Sample preparation: Test compounds were diluted in assay buffer (imidazole, glycerol-2-phosphate, EGTA, MnCl ₂ , MgCl ₂ , BSA) in semi-log increments.

2) Bead preparation

a.) Rinse the beads by centrifugation at 500 g

b.) Reducing the beads with PBS and EDTA to produce 20% bead slurry

3) Pre-incubation of the reaction mixture without the substrate (assay buffer, DTT, ATP, ³³ P ATP) and the substrate at 30 ° C (assay buffer, DTT, ATP, ³³ P ATP, peptide substrate) Mix for 15 minutes.

4) For the initial analysis, 10 μL of Btk-containing enzyme buffer (imidazole, glycerol-2-phosphorus) The acid ester, BSA) and 10 μL of the test compound were pre-incubated for 10 minutes at room temperature.

5) Add 30 μL of the reaction mixture containing or containing the substrate to Btk and the compound.

6) Incubate 50 μL of the entire assay mixture for 30 minutes at 30 °C.

7) Transfer 40 μL of the analyte to 150 μL of the bead slurry in the filter tray to stop the reaction.

8) Wash the filter disc after 30 minutes by the following steps

a. 3 × 250 μL NaCl

b. 3 × 250 μL NaCl (containing 1% phosphoric acid)

_{c. 1 × 250 μL H 2} O

9) Dry the tray at 65 ° C for 1 hour or dry at room temperature overnight

10) Add 50 μL of microscint-20 and count ³³ P cpm with a scintillation counter.

Calculate the percentage of activity from the original data in cpm

Percentage of activity = (sample - bkg) / (total activity - bkg) × 100

IC ₅₀ calculated from percent activity using a single site dose response sigmoidal model

y=A+((BA)/(1+((x/C) ^D )))

x = compound concentration, y = activity %, A = minimum, B = maximum, C = IC ₅₀ , D = 1 (hill slope)

Inhibition of B cell activation in whole blood measured by CD69 expression

The procedure for testing the ability of Btk inhibitors to inhibit B cell receptor-mediated B cell activation in human blood is as follows: Human whole blood (HWB) is obtained from healthy volunteers with the following restrictions: 24 hours without medication, non-smoking By. Blood was collected by venipuncture into a Vacutainer tube in anticoagulation with sodium heparin. Test compounds were diluted in PBS (20x) to ten times the desired starting drug concentration, followed by three-fold serial dilutions in PBS containing 10% DMSO to produce a nine point dose response curve. 5.5 μl of each compound dilution was added to a 2 ml 96-well V-type chassis (Analytical Sales and Services, #59623-23); 5.5 μl of PBS containing 10% DMSO was added to the control wells and non-irritant wells. HWB (100 μl) was added to each well and after mixing, the disk was incubated at 37 ° C, 5% CO ₂ , 100% humidity for 30 minutes. Add goat F(ab')2 anti-human IgM (Southern Biotech, #2022-14) (10 μl 500 μg/ml solution, 50 μg/ml final concentration) to each well (except for non-irritant wells) with mixing ) and cultivating the tray for another 20 hours.

At the end of the 20-hour incubation, the sample was labeled with a fluorescent probe (15 μl PE mouse anti-human CD20, BD Pharmingen, #555623, and/or 20 μl APC mouse anti-human CD69, BD Pharmingen, #555533 ) Incubate together for 30 minutes at 37 ° C, 5% CO ₂ , 100% humidity. Induced controls, unstained and single stains were included to compensate for the adjustment and initial voltage settings. The sample was then dissolved in 1 ml of 1X Pharmingen Lysis Buffer (BD Pharmingen #555899) and the plate was centrifuged at 1800 rpm for 5 minutes. The supernatant was removed via aspiration and the remaining pellet was dissolved again with another 1 ml of 1X Pharmingen lysis buffer and the disk was centrifuged as previously described. The supernatant was aspirated and the remaining pellet was washed in FAC buffer (PBS + 1% FBS). After the final centrifugation, the supernatant was removed and the pellet was resuspended in 180 μl of FAC buffer. Samples were transferred to 96-well plates suitable for operation on the HTS 96-well system of the BD LSR II flow cytometer.

Data were obtained using the excitation and emission wavelengths appropriate for the fluorophore used and the percentage of positive cell values was obtained using Cell Quest software. The results were initially analyzed by FACS analysis software (Flow Jo). Of test compound IC ₅₀ defined as the CD69 positive cells (which is also anti-IgM stimulation after CD20-positive) percentage reduction of the concentration of 50% (average of eight control wells, the average of eight wells stimulated subtracts After the background of the object). Using the version 3 XLfit software, Equation 201 calculates the IC ₅₀ value.

Representative compound data for this assay are listed in Table II below.

Inhibition of B cell activation - B cell FLIPR assay in Ramos cells

The inhibition of B cell activation by the compounds of the invention is demonstrated by determining the effect of the test compound against IgM-stimulated B cell responses.

The B cell FLIPR assay is a cell-based functional assay that measures the effect of a potential inhibitor against intracellular calcium increase caused by IgM antibody stimulation. Ramos cells (human Burkitt's lymphoma cell line, ATCC No. CRL-1596) were cultured in growth medium (described below). On the day before the analysis, Ramos cells were resuspended in fresh growth medium (same as described above) and set to a concentration of 0.5 x 10 ⁶ per ml in tissue culture flasks. On the day of analysis, cells were counted and set in tissue culture flasks in growth medium supplemented with 1 μM FLUO-3AM (TefLabs Cat. No. 0116, prepared in anhydrous DMSO and 10% Pluronic acid). The concentration was 1 × 10 ⁶ /mL and incubated at 37 ° C (4% CO ₂ ) for 1 hour. To remove the extracellular dye, the cells were harvested by centrifugation (5 min, 1000 rpm), resuspended at 1 x 10 ⁶ cells/ml in FLIPR buffer (described below) and then at 1 x 10 ⁵ Cells/wells were dispensed into 96-well black/transparent disks (BD Cat. No. 356692) coated with poly-D-lysine. Test compounds were added at various concentrations ranging from 100 μM to 0.03 μM (7 concentrations, detailed below) and incubated with the cells for 30 minutes at room temperature. Ramos cell Ca ²⁺ signaling was stimulated by the addition of 10 μg/mL anti-IgM (Southern Biotech, Cat. No. 2020-01) and captured by FLIPR (Molecular Devices, CCD camera with argon laser at 480 nM excitation) Image of 96-well plate) measurement.

Medium / Buffer:

Growth medium: L-glutamic acid (Invitrogen, Cat. No. 61870-010), 10% fetal bovine serum (FBS, Summit Biotechnology catalog number FP-100-05), 1 mM sodium pyruvate (Invitrogen catalog number 11360- 070) RPMI 1640 medium.

FLIPR buffer: HBSS (Invitrogen, catalog number 141175-079), 2 mM CaCl ₂ (Sigma catalog number C-4901), HEPES (Invitrogen, catalog number 15630-080), 2.5 mM Probenecid (Sigma, Catalog No. P-8761), 0.1% BSA (Sigma, Cat. No. A-7906), 11 mM Glucose (Sigma, Cat. No. G-7528) Compound Dilution Details: To achieve a maximum final assay concentration of 100 μM, to 576 μL FLIPR 24 μL of 10 mM compound stock solution (prepared in DMSO) was added directly to the buffer. Test compounds were diluted in FLIPR buffer (using a Biomek 2000 robotic pipette) to produce the following dilution protocol: vehicle, 1.00 x 10 ^-4 M, 1.00 x 10 ^-5 , 3.16 x 10 ^-6 , 1.00 x 10 ^-6 , 3.16 × 10 ^-7 , 1.00 × 10 ^-7 , 3.16 × 10 ^-8 .

Analysis and analysis:

Intracellular calcium increase was reported using max-min statistic (using the Molecular Devices FLIPR control and statistical output software, subtracting the resting baseline from the peak produced by the addition of the stimulating antibody). Using a nonlinear curve fit (GraphPad Prism software) measured IC _50.

Mouse collagen-induced arthritis (mCIA)

On day 0, mice were injected with an emulsion of type II collagen in Complete Freund's adjuvant (CFA) at several points on the base or back of the tail (intradermal injection (i.d.)). After collagen immunization, the animals will develop arthritis in about 21 to 35 days. Systemic administration of Freund's incomplete adjuvant with collagen on day 21 (Incomplete Freund's adjuvant, IFA; intradermal injection) causes simultaneous onset of arthritis (enhanced immunity). Any episode of mild arthritis in animals was scored daily after day 20 (score 1 or 2; see score below description), which is a signal to enhance immunity. After booster immunization, the mice are scored and the candidate therapeutic agent is administered for a specified period of time (typically 2-3 weeks) and the frequency of dosing is once daily (QD) or twice daily (BID).

Rat collagen-induced arthritis (rCIA)

On day 0, emulsions of bovine type II collagen in incomplete Freund's adjuvant (IFA) were injected intradermally (i.d.) at several locations on the back of the rat. On about day 7, an injection of collagen emulsion (intradermal injection) was boosted at the base of the tail or at the back of the back. Arthritis is usually observed 12-14 days after the initial collagen injection. From the 14th day onwards, the development of arthritis in animals can be assessed as described below (arthritis assessment). The animal begins to administer the candidate therapeutic agent in a prophylactic manner on a secondary challenge for a specified period of time (typically 2-3 weeks) and is administered once daily (QD) or twice daily (BID).

Arthritis assessment:

In both models, a scoring system was used to quantify the development of inflammation of the paw and the amine joint, which involved assessing 4 paws in accordance with the criteria described below:

Rating: 1 = swelling of the paw or one of the toes and/or redness.

2 = Two or more joints are swollen.

3 = The whole paw is swollen and involves more than two joints.

4 = severe arthritis of the entire paw and toe.

A baseline measurement was performed on day 0 and started again on the first symptom or swelling, up to 3 times per week until the end of the experiment. The arthritis index of each mouse was obtained by adding 4 scores of individual paws, and each animal had a maximum score of 16.

Rat in vivo asthma model

Male Brown-Norway rats were sensitized intraperitoneally with 0.2 ml of alum (100 μg) of OA (ovalbumin) once a week for 3 weeks (Day 0, Day 7, and 14) day). On day 21 (one week after the last sensitization), rats were administered subcutaneously with vehicle or compound formulation 0.5 times before OA aerosol challenge (1% OA for 45 minutes) and after challenge 4 Hours or 24 hours. At the time of sacrifice, serum and plasma were collected from all animals for serology and PK, respectively. The tracheal cannula was inserted and the lungs were lavaged 3 times with PBS. The total white blood cell count and white blood cell classification count of BAL fluid were analyzed. The total number of white blood cells in a cell aliquot (20-100 μl) was determined by a Coulter counter. For white blood cell sorting, 50-200 μl samples were centrifuged in Cytospin and slides were stained with Diff-Quik. Monocytes, eosinophils, neutrophils, and lymphocytes were counted under light microscopy using standard morphological criteria and the ratios are expressed as a percentage. Representative Btk inhibitors showed a reduction in total white blood cell counts in BAL of OA sensitized and challenged rats compared to control levels.

The foregoing invention has been described in considerable detail for the purposes of clarity and understanding. It will be apparent to those skilled in the art that variations and modifications can be made within the scope of the appended claims. Therefore, the above description is to be considered as illustrative and not restrictive. Therefore, the scope of the invention should not be determined by the description of the invention, but the scope of the appended claims and the equivalents of the equivalents of the claims.

All of the patents, patent applications, and publications cited in this application are hereby incorporated by reference in their entirety for all purposes in the the the the the general.

Claims (20)

a compound of formula I, among them: Is a single bond or a double bond; X is CH, CH ₂ or N; R is H, -R ¹ , -R ¹ -R ² -R ³ , -R ¹ -R ³ or -R ² -R ³ ; R ¹ An aryl group, a heteroaryl group, a bicyclic heteroaryl group, a cycloalkyl group or a heterocycloalkyl group, each of which may optionally be subjected to one or more lower alkyl, hydroxy, hydroxy lower alkyl, lower carbon numbers Alkoxy, halo, nitro, amine, decyl, cyano, pendant or lower haloalkyl; R ² is -C(=O), -C(=O)O, -C(=O)NR ^2' , -NHC(=O)O, -C(R ^2' ) ₂ , -O, -S, -C(=NH)NR ^2' or -S(=O) ₂ Each R ^{2 ' is} independently H or a lower alkyl group; R ³ is H or R ⁴ ; R ⁴ is a lower alkyl group, a lower carbon haloalkyl group, a lower alkoxy group, an amine group, Lower alkylalkylamino, cycloalkylamino, lower dialkylamino, aryl, arylalkyl, alkylaryl, heteroaryl, alkylheteroaryl, heteroarylalkane Base, cycloalkyl, alkylcycloalkyl, cycloalkylalkyl, heterocycloalkyl, alkylheterocycloalkyl, heterocycloalkylalkyl, bicyclic cycloalkyl, bicycloheterocycloalkyl, spiro ring An alkyl group, a spiroheterocycloalkyl group or a bicyclospirocycloalkyl group, each of which is optionally one or more low C, alkyl, halo, lower alkylamino, lower dialkylamino, hydroxy, hydroxy lower alkyl, lower alkoxy, lower alkyl alkoxy, halo , nitro, amine, decyl, fluorenyl, cyano, pendant oxy, sulfonyl, lower alkyl sulfonyl, fluorenyl, hydroxyamino, carboxy, amine carbyl, amine a formate group, a halogen lower alkoxy group, a heterocycloalkyl group or a halogen lower alkyl group, wherein two lower alkyl groups may together form a ring; Y ⁴ is Y ^4a , Y ^4b , Y ^4c or Y ^4d ; Y ^4a is H or halogen; Y ^4b is a lower alkyl group, which is optionally selected from one or more selected from the group consisting of a lower halogen haloalkyl group, a halogen group, a hydroxyl group, an amine group, a cyano group and a lower group. Substituted by a substituent of the group of carbon number alkoxy groups; Y ^4c is a lower carbon cycloalkyl group, which is optionally selected from one or more selected from the group consisting of a lower alkyl group, a lower carbon haloalkyl group, a halogen group, a hydroxyl group, Substituted with a group of amine, cyano and lower alkoxy groups; and Y ^4d is an amine group, optionally via one or more lower alkyl, alkoxy lower alkyl or Hydroxylower alkyl substitution; or its medicinal Acceptable salt thereof. The compound of claim 1, wherein It is a double bond and X is N. The compound of claim 1, wherein It is a single bond and X is CH ₂ . The compound of any one of claims 1 to 3, wherein R is -R ¹ -R ² -R ³ ; R ¹ is pyridyl; R ² is -S(=O) ₂ , and R ³ is R ⁴ ; R ⁴ is a lower alkyl group. The compound of any one of claims 1 to 3, wherein R is -R ¹ -R ² -R ³ ; R ¹ is pyridyl; R ² is -C(CH ₃ ) ₂ ; R ³ is R ⁴ ; R ⁴ is a lower alkylamino group, a lower dialkylamino group or a heterocycloalkyl group optionally substituted with one or more lower alkyl groups. The compound of any one of claims 1 to 3, wherein R is -R ¹ -R ² -R ³ ; R ¹ is phenyl or pyridyl; R ² is -C(=O); R ³ is R ⁴ And R ⁴ is morpholine or piperazine, which is optionally substituted with one or more lower alkyl groups. The compound of claim 2, wherein Y ⁴ is a third butyl group. The compound of claim 3, wherein Is a double bond, X is CH, and Y ⁴ is Wherein Y ⁵ and Y ^{6 are} independently H, a lower alkyl group or a lower carbon haloalkyl group. The compound of claim 2, wherein Y ⁴ is Wherein Y ⁵ is H, halogen, lower alkyl or lower haloalkyl. The compound of claim 2, wherein Y ⁴ is Wherein Y ⁵ and Y ^{6 are} independently H or a lower alkyl group. The compound of claim 7, wherein R is -R ¹ -R ³ ; R ¹ is pyridyl or pyrazolopyrazine; R ³ is R ⁴ ; and R ⁴ is optionally substituted lower alkyl, Heterocycloalkyl or alkylheterocycloalkyl. The compound of claim 7, wherein R is -R ¹ -R ² -R ³ ; R ¹ is pyridyl; R ² is -C(CH ₃ ) ₂ ; R ³ is R ⁴ ; and R ⁴ is a low carbon number An alkylamino group, a lower dialkylamino group or a heterocycloalkyl group optionally substituted with one or more lower alkyl groups. The compound of claim 7, wherein R is -R ¹ -R ² -R ³ ; R ¹ is pyridyl; R ² is -C(=O); R ³ is R ⁴ ; and R ⁴ is optionally substituted Heterocycloalkyl or bicyclospirocycloalkyl. The compound of claim 13, wherein R ⁴ is optionally substituted morpholine or piperazine. The compound according to any one of claims 1 to 3 and 7 to 14, which is selected from the group consisting of 2-(3-{5-[5-(2-azetidin-1-yl-)- 1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl}-2- Hydroxymethyl-phenyl)-6-tert-butyl-2H-phthalazin-1-one; 6-Tertibutyl-2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1',2',3',4',5',6' -hexahydro-[3,4']bipyridin-6-ylamino)-6-oxooxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-2H-pyridazine- 1-keto; 2-{3-[5-(5-azetidin-1-ylmethyl-1-methyl-1H-pyrazol-3-ylamino)-1-methyl-6 -Sideoxy-1,6-dihydro-pyridazin-3-yl]-2-hydroxymethyl-phenyl}-6-tert-butyl-2H-phthalazin-1-one; 6-third Butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5-((S)-1-methyl-pyrrolidin-2-yl)-pyridin-2-ylamino) -6-Sideoxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one; 6-t-butyl-2-(2-hydroxyl) 3-(1-methyl-5-[5-((S)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-6-oxo-oxyl-1, 6-dihydro-pyridazin-3-yl}-phenyl)-2H-phthalazin-1-one; 6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl- 5-[5-((R)-1-methyl-pyrrolidin-3-yl)-pyridin-2-ylamino]-6-oxooxy-1,6-dihydro-pyridazine-3- }--phenyl)-2H-phthalazin-1-one; 2-(2-{2-hydroxymethyl-3-[1-methyl-5-(1'-methyl-1',2' ,3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-o-oxy-1,6-dihydro-pyridazine-3- Base]-phenyl}-1-sidedoxy-1,2 -dihydro-pyridazin-6-yl)-2-methyl-propionitrile; 6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(5-A 4-,5,6,7-tetrahydro-pyrazolo[1,5-a]pyrazin-2-ylamino)-6-o-oxy-1,6-dihydro-pyridazine-3 -yl]-phenyl}-2H-phthalazin-1-one; 6-t-butyl-2-{3-[5-(1'-ethyl-1',2',3',4' ,5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-1-methyl-6-oxirane-1,6-dihydro-pyridazin-3-yl ]-2-hydroxymethyl-phenyl}-2H-phthalazin-1-one; 6-t-butyl-2-(2-hydroxymethyl-3-{1-methyl-5-[5- (morpholine-4-carbonyl)-pyridin-2-ylamino]-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-phenyl)-2H-pyridazine-1- ketone; 2-[2-(3-{5-[5-(2-azetidin-1-yl-1,1-dimethyl-ethoxy)-pyridin-2-ylamino]-1 -methyl-6-o-oxy-1,6-dihydro-pyridazin-3-yl}-2-hydroxymethyl-phenyl)-1-yloxy-1,2-dihydro-pyridazine -6-yl]-2-methyl-propionitrile; 6-t-butyl-2-[2-hydroxymethyl-3-(5-{5-[2-(3-hydroxy-propylamino) )-1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-oxirane-1,6-dihydro-pyridazin-3-yl)- Phenyl]-2H-phthalazin-1-one; 6-t-butyl-2-{2-hydroxymethyl-3-[1-methyl-5-(1'-oxetan-3 -yl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6-oxirane-1,6- Dihydro-pyridazin-3-yl]-phenyl}-2H-phthalazin-1-one; 6-t-butyl-2-[3-(5-{5-[2-(3,3- Difluoro-azetidin-1-yl)-1,1-dimethyl-ethoxy]-pyridin-2-ylamino}-1-methyl-6-sideoxy-1,6 -dihydro-pyridazin-3-yl)-2-hydroxymethyl-phenyl]-2H-pyridazin-1-one; and 2-(2-{2-hydroxymethyl-3-[1-- 5-(1'-methyl-1',2',3',4',5',6'-hexahydro-[3,4']bipyridin-6-ylamino)-6- Sideoxy-1,6-dihydro-pyridazin-3-yl]-phenyl}-1-sidedoxy-1,2,3,4-tetrahydro-isoquinolin-6-yl)-2 -Methyl-propionitrile. A pharmaceutical composition comprising a mixture of a compound according to any one of claims 1 to 15 and at least one pharmaceutically acceptable carrier, excipient or diluent. Use of a compound according to any one of claims 1 to 15 for the manufacture of a medicament for the treatment of an inflammatory or autoimmune disorder. The use of claim 17, wherein the condition is rheumatoid arthritis or asthma. A compound according to any one of claims 1 to 3 and 7 to 14 for use in the treatment of an inflammatory or autoimmune disorder. The compound of claim 19, wherein the condition is rheumatoid arthritis or asthma.